Vaccines for coronavirus and methods of using the same

ABSTRACT

Disclosed are compositions comprising an expressible nucleic acid sequence comprising a first nucleic acid sequence comprising a sequence that encodes a self-assembling polypeptide or a pharmaceutically acceptable salt thereof and a second nucleic acid sequence comprising a sequence that encodes an antigen from a virus from the family of Coronaviridae. In some embodiments, the expressible nucleic acid sequence further comprises a nucleic acid sequence encoding at least one viral antigen or a pharmaceutically acceptable salt thereof. In some embodiments, the expressible nucleic acid sequence further comprises at least one nucleic acid sequence encoding a linker.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.63/008,733 filed Apr. 11, 2020, the entire content of which is herebyincorporated by reference in its entirety into the present application.

SUBMISSION OF SEQUENCE LISTING

The Sequence Listing associated with this application is filed inelectronic format as a text file and hereby incorporated by referenceinto the specification in its entirety. The name of the text filecontaining the Sequence Listing is 37925_0008_SL_ST25 and the size ofthe text file is 675 KB.

BACKGROUND

Vaccination is an extremely important public health measure that hasdemonstrated prophylactic and therapeutic utility against manyinfectious diseases^([1-3]), and impacted some forms of cancer^([4]). Inthe past decade, advances in material engineering has allowed for thedevelopment and study of a new generation of vaccines, such asnanoparticle vaccines^([5-7]). Hepatitis B and human papillomavirus(HPV) vaccines are examples of such self-assembling virus-like particleswhich have impacted millions of people^([8, 9]). Nanoparticles may comein several shapes and forms. Inorganic materials^([10, 11]), nontoxicphospholipids^([12]), virus-like particles (VLPs) or self-assemblingprotein nanoparticles (SAPN)^([13-16]) can all scaffold and presentantigens in repetitive multimeric manners to robustly stimulate immunityin animal models^([16-18]). An exemplary disclosure on compositionscomprising such self-assembling vaccines and methods of preparing andusing the same was provided in PCT application No. PCT/US2019/68444filed on Dec. 23, 2019 based on U.S. Provisional Application No.62/784,318 filed Dec. 21, 2018, each of which is incorporated byreference in its entirety.

Another example of such new generation of vaccines are recombinantnative-like viral trimer vaccines^([70-72]). An exemplary disclosure oncompositions comprising nucleic acids encoding such structural trimersand methods of preparing and using the same was provided in PCTapplication No. PCT/US2020/26948 filed on Apr. 6, 2020 based on U.S.Provisional Application No. 62/829,629 filed on Apr. 4, 2019, each ofwhich is incorporated by reference in its entirety.

Coronaviruses are a group of viruses that cause diseases in mammals andbirds. Coronaviruses were first discovered in the 1960s. The earliestvirus from the family of Coronaviridae discovered were infectiousbronchitis virus in chickens and two viruses from the nasal cavities ofhuman patients with the common cold that were subsequently named humancoronavirus 229E (HCoV-229E) and human coronavirus OC43 (HCoV-OC43).Other members of this family have since been identified, includingSARS-CoV in 2003, HCoV NL63 in 2004, HKU1 in 2005, MERS-CoV in 2012, andSARS-CoV-2 (formerly known as 2019-nCoV or novel coronavirus 2019, whichcaused the global COVID-19 pandemic). Most of these have involvedserious respiratory tract infections.

Symptoms of coronavirus infection vary in species. In chicken, forinstance, they cause an upper respiratory tract disease, while in cowsand pigs they cause diarrhea. In humans, coronaviruses cause respiratorytract infections that are typically mild, such as some cases of thecommon cold (among other possible causes, predominantly rhinoviruses),though rarer forms can be lethal, such as SARS, MERS, and COVID-19.

Coronaviruses vary significantly in risk factor. Some can kill more than30% of those infected (such as MERS-CoV), and some are relativelyharmless, such as the common cold. Coronaviruses cause flu-likesymptoms, such as fever and sore throat from swollen adenoids, primarilyin the winter and early spring seasons. Coronaviruses can causepneumonia—either direct viral pneumonia or a secondary bacterialpneumonia—and may cause bronchitis—either direct viral bronchitis or asecondary bacterial bronchitis. The much publicized human coronavirusdiscovered in 2003, SARS-CoV, which causes severe acute respiratorysyndrome (SARS), has a unique pathogenesis because it causes both upperand lower respiratory tract infections. The novel human coronavirusdiscovered in 2019, SARS-CoV-2, causes mild symptoms, most often fever,dry cough, and shortness of breath, to complications including pneumoniaand acute respiratory distress syndrome.

SUMMARY OF EMBODIMENTS

Since it was emerged in late 2019, SARS-CoV-2 was rapidly characterizedas a new member of the betacoronavirus genus, closely related to severalbat coronaviruses and to severe acute respiratory syndrome coronavirus(SARS-CoV). Compared to SARS-CoV, however, SARS-CoV-2 appears to be morereadily transmitted from human to human.

SARS-CoV-2 makes use of a densely glycosylated Spike (S) protein to gainentry into host cells. The S protein is a trimeric class I fusionprotein that exists in a metastable prefusion conformation thatundergoes a substantial structural rearrangement to fuse the viralmembrane with the host cell membrane^([73, 74]). This process istriggered when the S1 subunit of the S protein binds to a host cellreceptor. Receptor binding destabilizes the prefusion trimer, resultingin shedding of the S1 subunit and transition of the S2 subunit to astable postfusion conformation^([75]). To engage a host cell receptor,the receptor-binding domain (RBD) of S1 undergoes hinge-likeconformational movements that transiently hide or expose thedeterminants of receptor binding. These two states are referred to asthe “down” conformation and the “up” conformation, where downcorresponds to the receptor-inaccessible state and up corresponds to thereceptor-accessible state, which is thought to be lessstabler^([76-79]). Because of the indispensable function of the Sprotein, it represents a target for vaccine design and development. SeeWrapp et al., Science, 2020, 367, 1260-1263.

In one aspect, the disclosure relates to a composition comprising anexpressible nucleic acid sequence. In some embodiments, the expressiblenucleic acid sequence comprises a first nucleic acid sequence encoding ascaffold domain comprising a self-assembling polypeptide and a secondnucleic acid sequence encoding a viral antigen from a virus of thefamily Coronaviridae. In some embodiments, the expressible nucleic acidsequence further comprises a third nucleic acid sequence encoding aleader sequence. In some embodiments, the expressible nucleic acidsequence further comprises a nucleic acid sequence encoding a linker,said nucleic acid sequence positioned between the first nucleic acidsequence and the second nucleic acid sequence in the 5′ to 3′orientation.

In other embodiments, the expressible nucleic acid sequence comprises afirst nucleic acid sequence encoding a leader sequence and a secondnucleic acid sequence encoding a soluble viral trimer or a solublemonomer thereof, wherein the soluble viral trimer or the soluble monomerthereof is from a virus of the family Coronaviridae. In someembodiments, the expressible nucleic acid sequence further comprises athird nucleic acid sequence encoding a linker. In some embodiments, thesoluble viral trimer or the soluble monomer thereof comprises at least aportion of SARS-CoV-2 spike protein. In some embodiments, theexpressible nucleic acid sequence further comprises a nucleic acidsequence encoding a self-assembling polypeptide or a functional fragmentor variant thereof.

In some embodiments, the disclosure relates to a vaccine comprising apolypeptide. In some embodiments, the polypeptide comprises a scaffolddomain comprising a self-assembling polypeptide and an antigen domaincomprising a viral antigen from a virus of the family Coronaviridae. Insome embodiments, the polypeptide further comprises a leader sequence.In some embodiments, the polypeptide further comprises a linker domaincomprising a linker peptide located between the scaffold domain and theantigen domain. In some embodiments, the polypeptide comprises at least70% sequence identity to SEQ ID NO: 70, SEQ ID NO: 73, SEQ ID NO: 76,SEQ ID NO: 79, SEQ ID NO: 82, SEQ ID NO: 85, SEQ ID NO: 88 or SEQ ID NO:91, or a functional fragment or variant thereof.

In some embodiments, the polypeptide comprises a leader sequence and anantigen domain comprising a soluble viral trimer or a soluble monomerthereof, wherein the soluble viral trimer or the soluble monomer thereofis from a virus of the family Coronaviridae. In some embodiments, thepolypeptide further comprises one or a plurality of linker. In someembodiments, the soluble viral trimer or the soluble monomer thereofcomprises at least a portion of SARS-CoV-2 spike protein. In someembodiments, the polypeptide further comprises a self-assemblingpolypeptide or a functional fragment or variant thereof. In someembodiments, the polypeptide comprises at least 70% sequence identity toSEQ ID NO: 94, SEQ ID NO: 97, SEQ ID NO: 100, SEQ ID NO: 103, SEQ ID NO:106, SEQ ID NO: 109, SEQ ID NO: 112, SEQ ID NO: 115, SEQ ID NO: 118, SEQID NO: 121, SEQ ID NO: 124, SEQ ID NO: 127, SEQ ID NO: 130, SEQ ID NO:133, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 142, SEQ ID NO: 145, SEQID NO: 148, SEQ ID NO: 151, SEQ ID NO: 154, SEQ ID NO: 157 or SEQ ID NO:160, or a functional fragment or variant thereof.

In a further aspect, the disclosure relates to a cell comprising anexpressible nucleic acid sequence. In some embodiments, the expressiblenucleic acid sequence comprises a first nucleic acid sequence encoding ascaffold domain comprising a self-assembling polypeptide and a secondnucleic acid sequence encoding a viral antigen from a virus of thefamily Coronaviridae. In some embodiments, the expressible nucleic acidsequence further comprises a third nucleic acid sequence encoding aleader sequence. In some embodiments, the expressible nucleic acidsequence further comprises a nucleic acid sequence encoding a linker,said nucleic acid sequence positioned between the first nucleic acidsequence and the second nucleic acid sequence in the 5′ to 3′orientation. In some embodiments, the expressible nucleic acid sequencecomprises a nucleic acid sequence encoding a leader sequence, a nucleicacid sequence encoding a scaffold domain, a nucleic acid sequenceencoding a linker, and a nucleic acid sequence encoding a viral antigenfrom Coronaviridae, in the 5′ to 3′ orientation.

In some embodiments, the expressible nucleic acid sequence comprises afirst nucleic acid sequence encoding a leader sequence and a secondnucleic acid sequence encoding a soluble viral trimer or a solublemonomer thereof, wherein the soluble viral trimer or the soluble monomerthereof is from a virus of the family Coronaviridae. In someembodiments, the expressible nucleic acid sequence further comprises athird nucleic acid sequence encoding a linker. In some embodiments, thesoluble viral trimer or the soluble monomer thereof comprises at least aportion of SARS-CoV-2 spike protein. In some embodiments, theexpressible nucleic acid sequence further comprises a nucleic acidsequence encoding a self-assembling polypeptide or a functional fragmentor variant thereof.

In some embodiments, the self-assembling polypeptide of the disclosureis from Aquifex aeolicus, Helicobacter pylori, Pyrococcus furiosus orThermotoga maritima. In some embodiments, the self-assemblingpolypeptide of the disclosure comprises at least 70% sequence identityto SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO:16, SEQ ID NO: 18 or SEQ ID NO: 20, or a functional fragment or variantthereof.

In some embodiments, the viral antigen of the disclosure is an antigenfrom a coronavirus. In some embodiments, the viral antigen is an antigenfrom SARS-CoV-2. In some embodiments, the viral antigen comprises atleast 70% sequence identity to SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO:64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 or SEQ ID NO: 177, or afunctional fragment or variant thereof.

In some embodiments, the leader sequence of the disclosure comprises atleast 70% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 5, or afunctional fragment or variant thereof.

In some embodiments, the linker of the disclosure comprises at least 70%sequence identity to SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ IDNO: 25, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46,SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO:56 or SEQ ID NO: 58, or a functional fragment or variant thereof.

In some embodiments, the expressible nucleic acid sequence of thedisclosure comprises at least 70% sequence identity to SEQ ID NO: 68,SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO:75, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 81, SEQ IDNO: 83, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 89 orSEQ ID NO: 90, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence of the disclosureencodes a polypeptide comprising at least 70% sequence identity to SEQID NO: 70, SEQ ID NO: 73, SEQ ID NO: 76, SEQ ID NO: 79, SEQ ID NO: 82,SEQ ID NO: 85, SEQ ID NO: 88 or SEQ ID NO: 91, or a functional fragmentor variant thereof. In some embodiments, the expressible nucleic acidsequence of the disclosure comprises at least 70% sequence identity toSEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO:98, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 104, SEQID NO: 105, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO:111, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 117, SEQID NO: 119, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO:125, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 131, SEQID NO: 132, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO:138, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 144, SEQID NO: 146, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO:152, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 158 orSEQ ID NO: 159, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence of the disclosureencodes a polypeptide comprising at least 70% sequence identity to SEQID NO: 94, SEQ ID NO: 97, SEQ ID NO: 100, SEQ ID NO: 103, SEQ ID NO:106, SEQ ID NO: 109, SEQ ID NO: 112, SEQ ID NO: 115, SEQ ID NO: 118, SEQID NO: 121, SEQ ID NO: 124, SEQ ID NO: 127, SEQ ID NO: 130, SEQ ID NO:133, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 142, SEQ ID NO: 145, SEQID NO: 148, SEQ ID NO: 151, SEQ ID NO: 154, SEQ ID NO: 157 or SEQ ID NO:160, or a functional fragment or variant thereof.

In some embodiments, the expressible nucleic acid sequence of thedisclosure is operably linked to one or a plurality of regulatorysequences. In some embodiments, the expressible nucleic acid sequence ofthe disclosure is comprised in a nucleic acid molecule. In someembodiments, the expressible nucleic acid sequence of the disclosure iscomprised in a nucleic acid molecule which is a plasmid.

In some embodiments, the disclosure relates to a pharmaceuticalcomposition comprising a pharmaceutically effective amount of any of thecomposition disclosed herein and a pharmaceutically acceptable carrier.In some embodiments, the pharmaceutical composition comprises any of thedisclosed composition from about 1 to about 100 micrograms. In someembodiments, the pharmaceutical composition comprises any of thedisclosed composition from about 1 to about 20 micrograms.

In some embodiments, the disclosure relates to a method of vaccinating asubject comprising administering a therapeutically effective amount ofany of the disclosed pharmaceutical composition to the subject.

In other aspects, the disclosure relates to a method of inducing animmune response in a subject comprising administering to the subject anyof the disclosed pharmaceutical composition. In some embodiments, theimmune response is an antigen-specific immune response againstSARS-CoV-2. In some embodiments, the subject is diagnosed with orsuspected of having a SARS-CoV-2 infection. In some embodiments, theimmune response is an antigen-specific immune response against aSARS-CoV-2 antigen. In some embodiments, the immune response is anantigen-specific immune response against a SARS-CoV-2 spike antigen.

In some embodiments, the disclosure relates to a method of neutralizingone or a plurality of viruses in a subject comprising administering tothe subject any of the disclosed pharmaceutical composition.

In some embodiments, the disclosure relates to a method of stimulating atherapeutically effective antigen-specific immune response against avirus in a mammal infected with a virus comprising administering atherapeutically effective amount of any of the disclosed pharmaceuticalcomposition. In some embodiments, the subject is infected withSARS-CoV-2.

In some embodiments, the disclosure relates to a method of inducingexpression of a self-assembling vaccine in a subject comprisingadministering any of the disclosed pharmaceutical composition. In someembodiments, the method is free of administering any polypeptidedirectly to the subject.

In some embodiments, the administering in any of the disclosed methodsis accomplished by oral administration, parenteral administration,sublingual administration, transdermal administration, rectaladministration, transmucosal administration, topical administration,inhalation, buccal administration, intrapleural administration,intravenous administration, intraarterial administration,intraperitoneal administration, subcutaneous administration,intramuscular administration, intranasal administration, intrathecaladministration, and intraarticular administration, or a combinationthereof. In some embodiments, the therapeutically effective amount usedin any of the disclosed methods is from about 1 to about 2000 microgramsof the expressible nucleic acid sequence. In some embodiments, thetherapeutically effective amount used in any of the disclosed methods isfrom about 1 to about 30 micrograms of the expressible nucleic acidsequence. In some embodiments, the disclosed method is free ofactivating any mannose-binding lectin or complement process. In someembodiments, the subject is a human. In some embodiments, thetherapeutically effective dose used in any of the disclosed method isfrom about 0.3 microgram of the composition per kilogram of the subjectto about 30 micrograms per kilogram of the subject. In some embodiments,the therapeutically effective dose used in any of the disclosed methodis from about 0.001 microgram of composition per kilogram of the subjectto about 0.05 microgram per kilogram of the subject.

The disclosure further relates to a DNA vaccine comprising anexpressible nucleic acid sequence encoding a polypeptide comprising atleast about 70% sequence identity to SEQ ID NO: 70, SEQ ID NO: 73, SEQID NO: 76, SEQ ID NO: 79, SEQ ID NO: 82, SEQ ID NO: 85, SEQ ID NO: 88 orSEQ ID NO: 91, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence of the disclosed DNAvaccine comprises at least about 70% sequence identity to SEQ ID NO: 68,SEQ ID NO: 71, SEQ ID NO: 74, SEQ ID NO: 77, SEQ ID NO: 80, SEQ ID NO:83, SEQ ID NO: 86, SEQ ID NO: 89, SEQ ID NO: 92, SEQ ID NO: 95, SEQ IDNO: 98, SEQ ID NO: 101, SEQ ID NO: 104, SEQ ID NO: 107, SEQ ID NO: 110,SEQ ID NO: 113, SEQ ID NO: 116, SEQ ID NO: 119, SEQ ID NO: 122, SEQ IDNO: 125, SEQ ID NO: 128, SEQ ID NO: 131, SEQ ID NO: 134, SEQ ID NO: 137,SEQ ID NO: 140, SEQ ID NO: 143, SEQ ID NO: 146, SEQ ID NO: 149, SEQ IDNO: 152, SEQ ID NO: 155 or SEQ ID NO: 158, or a functional fragment orvariant thereof. In some embodiments, the disclosed DNA vaccine furthercomprises a pharmaceutically acceptable excipient. In some embodiments,the pharmaceutically acceptable excipient further comprises an adjuvant.

The disclosure also relates to a RNA vaccine comprising an expressiblenucleic acid sequence encoding a polypeptide comprising at least about70% sequence identity to SEQ ID NO: 70, SEQ ID NO: 73, SEQ ID NO: 76,SEQ ID NO: 79, SEQ ID NO: 82, SEQ ID NO: 85, SEQ ID NO: 88 or SEQ ID NO:91, or a functional fragment or variant thereof. In some embodiments,the expressible nucleic acid sequence of the disclosed RNA vaccinecomprises at least about 70% sequence identity to SEQ ID NO: 69, SEQ IDNO: 72, SEQ ID NO: 75, SEQ ID NO: 78, SEQ ID NO: 81, SEQ ID NO: 84, SEQID NO: 87, SEQ ID NO: 90, SEQ ID NO: 93, SEQ ID NO: 96, SEQ ID NO: 99,SEQ ID NO: 102, SEQ ID NO: 105, SEQ ID NO: 108, SEQ ID NO: 111, SEQ IDNO: 114, SEQ ID NO: 117, SEQ ID NO: 120, SEQ ID NO: 123, SEQ ID NO: 126,SEQ ID NO: 129, SEQ ID NO: 132, SEQ ID NO: 135, SEQ ID NO: 138, SEQ IDNO: 141, SEQ ID NO: 144, SEQ ID NO: 147, SEQ ID NO: 150, SEQ ID NO: 153,SEQ ID NO: 156 or SEQ ID NO: 159, or a functional fragment or variantthereof. In some embodiments, the disclosed RNA vaccine furthercomprises a pharmaceutically acceptable excipient. In some embodiments,the pharmaceutically acceptable excipient further comprises an adjuvant.

The disclosure relates to a viral particle or a self-assemblyingparticle comprising either a pharmaceutically effective amount ofantigen from Coronaviridae in a trimer configuration and/or a viralparticle comprising a pharmaceutically effective amount of any nuclecacid sequence disclosed herein. In some embodiments, the viral particleis an adeno-associated vector (AAV) or lentiviral vector.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated in and constitute apart of this specification, illustrate several embodiments of thedisclosed method and compositions and together with the description,serve to explain the principles of the disclosed method andcompositions.

FIG. 1A-1B show the structure of SARS-CoV-2 in the prefusionconformation. FIG. 1A is a schematic of SARS-CoV-2 S primary structuregrayscaled by domain. Domains that could not be visualized in the finalmap are colored white. SS: signal sequence; S2′: S2′ protease cleavagesite; FP: fusion peptide; HR1: heptad repeat 1; CH: central helix; CD:connector domain; HR2: heptad repeat 2; TM: transmembrane domain; CT:cytoplasmic tail. Arrows denote protease cleavage sites. FIG. 1B showsthe side and top views of the prefusion structure of the SARS-CoV-2protein with a single RBD in the up conformation. The two RBD downprotomers are shown as cryo-EM density in either white or gray and theRBD up protomer is shown in ribbons grayscaled corresponding to theschematic in (A). Source: Wrapp et al., Science, 2020, 367, 1260-1263.

FIG. 2 shows that the fusion peptide from SARS-CoV2 was scaffolded ontoa modified version of Lumazine Synthase (60mer) with an expressiondomain as a genetic fusion. SARS-CoV-2 fusion peptide nanoparticle shownon the left is decorated by sixty copies of the fusion peptide.

FIG. 3A-3B depict the expression of 2 SARS-CoV-2 fusion peptidenanoparticle designs. FIG. 3A depicts the expression of constructWuhanS_FP_L9GT60_pVax. FIG. 3B depicts the expression of constructWuhanS_FP12_L9GT60_pVax. For SARS-Cov2 fusion peptide purification, DNAwas maxiprepped and transfected into Expi293 cells with PEI. Constructswere harvested on day 6. Constructs were purified by lectinchromatography and run on a size-exclusion column (GE Superdex S6increase). Control nanoparticles elute at ˜12.5 ml using the sameprotocols.

FIG. 4A-4E show the results of SARS-CoV-2 fusion peptide binding ELISA.FIG. 4A: Week 1 post vaccination; FIG. 4B: Week 2 post vaccination; FIG.4C: Week 3 post vaccination; FIG. 4D: Week 4 post vaccination; FIG. 4E:Comparison of endpoint titers to WuFuse1 between WuhanS_FP12_L9GT60_pVax(“S_FP12_L9GT60”) and WuhanS_FP_L9GT60_pVax (“S_FP_L9GT60”). ELISAplates were coated with 2 μg/mL streptavidin at room temperature for 8hours and then blocked overnight at 4° C. The next day, the plates wereincubated with fusion peptide at 2 μg/mL at room temp for 2 hours andthen with sera at 37° C. for 2 hours, followed by addition of anti-mouseIgG (H+L)-HRP to the plates. The plates were developed with TMBsubstrates for 5 minutes, absorbances at 450 nm and 570 nm wererecorded.

FIG. 5A-5E show the results of SARS-CoV-2 full-length spike proteinbinding ELISA. FIG. 5A: Week 1 post vaccination; FIG. 5B: Week 2 postvaccination; FIG. 5C: Week 3 post vaccination; FIG. 5D: Week 4 postvaccination; FIG. 5E: Comparison of endpoint titers to SARS-CoV-2full-length spike protein (“FL SARS2 Spike”) betweenWuhanS_FP12_L9GT60_pVax (“S_FP12_L9GT60”) and WuhanS_FP_L9GT60_pVax(“S_FP_L9GT60”). ELISA plates were coated with 1 μg/mL anti-histineantibody at room temp for 8 hours and then blocked overnight at 4° C.The next day, the plates were incubated with His-tagged SARS-CoV-2 spikeprotein at 2 μg/mL at room temperature for 2 hours and then with sera at37° C. for 2 hours, followed by addition of anti-mouse IgG (H+L)-HRP tothe plates. The plates were developed with TMB substrates for 5 minutes,absorbances at 450 nm and 570 nm were recorded.

DETAILED DESCRIPTION

The disclosed method and compositions may be understood more readily byreference to the following detailed description of particularembodiments and the examples included therein and to the figures andtheir previous and following description. It is to be understood thatthe disclosed method and compositions are not limited to specificsynthetic methods, specific analytical techniques, or to particularreagents unless otherwise specified, and, as such, may vary. It is alsoto be understood that the terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to limit thescope of the present disclosure which will be limited only by theappended claims.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the disclosure pertains.

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an,” and “the” include plural reference unless thecontext clearly dictates otherwise. Thus, for example, reference to “anucleic acid sequence” includes a plurality of nucleotides that areformed, reference to “the nucleic acid sequence” is a reference to oneor more nucleic acid sequences and equivalents thereof known to thoseskilled in the art, and so forth.

Ranges may be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, also specifically contemplated and considered disclosed isthe range from the one particular value and/or to the other particularvalue unless the context specifically indicates otherwise. Similarly,when values are expressed as approximations, by use of the antecedent“about,” it will be understood that the particular value forms another,specifically contemplated embodiment that should be considered disclosedunless the context specifically indicates otherwise. It will be furtherunderstood that the endpoints of each of the ranges are significant bothin relation to the other endpoint, and independently of the otherendpoint unless the context specifically indicates otherwise. The term“about” as used herein when referring to a measurable value such as anamount, a temporal duration, and the like, is meant to encompassvariations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or ±0.1% from the specifiedvalue, as such variations are appropriate to perform the disclosedmethods.

As used herein, the terms “activate,” “stimulate,” “enhance” “increase”and/or “induce” (and like terms) are used interchangeably to generallyrefer to the act of improving or increasing, either directly orindirectly, a concentration, level, function, activity, or behaviorrelative to the natural, expected, or average, or relative to a controlcondition. “Activate” in context of an immunotherapy refers to a primaryresponse induced by ligation of a cell surface moiety. For example, inthe context of receptors, such stimulation entails the ligation of areceptor and a subsequent signal transduction event. Further, thestimulation event may activate a cell and upregulate or downregulateexpression or secretion of a molecule. Thus, indirect or direct ligationof cell surface moieties, even in the absence of a direct signaltransduction event, may result in the reorganization of cytoskeletalstructures, or in the coalescing of cell surface moieties, each of whichcould serve to enhance, modify, or alter subsequent cellular responses.As used herein, the terms “activating CD8+ T cells” or “CD8+ T cellactivation” refer to a process (e.g., a signaling event) causing orresulting in one or more cellular responses of a CD8+ T cell (CTL),selected from: proliferation, differentiation, cytokine secretion,cytotoxic effector molecule release, cytotoxic activity, and expressionof activation markers. As used herein, an “activated CD8+ T cell” refersto a CD8+ T cell that has received an activating signal, and thusdemonstrates one or more cellular responses, selected fromproliferation, differentiation, cytokine secretion, cytotoxic effectormolecule release, cytotoxic activity, and expression of activationmarkers. Suitable assays to measure CD8+ T cell activation are known inthe art and are described herein.

The term “combination therapy” as used herein is meant to refer toadministration of one or more therapeutic agents in a sequential manner,that is, wherein each therapeutic agent is administered at a differenttime; as well as administration of these therapeutic agents, or at leasttwo of the therapeutic agents, in a substantially simultaneous manner.In some embodiments, the therapeutic agents are administered within 1,2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes within each other. Substantiallysimultaneous administration can be accomplished, for example, byadministering to the subject a single dose having a fixed ratio of eachtherapeutic agent or in multiple, individual doses for each of thetherapeutic agents. For example, one combination of the presentdisclosure may comprise a pooled sample of one or more nucleic acidmolecules comprising one or a plurality of expressible nucleic acidsequences and an adjuvant and/or an anti-viral agent administered at thesame or different times. In some embodiments, the pharmaceuticalcomposition of the disclosure can be formulated as a single,co-formulated pharmaceutical composition comprising one or more nucleicacid molecules comprising one or a plurality of expressible nucleic acidsequences and one or more adjuvants and/or one or more anti-viralagents. As another example, a combination of the present disclosure(e.g., DNA or RNA vaccines and anti-viral agent) may be formulated asseparate pharmaceutical compositions that can be administered at thesame or different time. As used herein, the term “simultaneously” ismeant to refer to administration of one or more agents at the same time.For example, in certain embodiments, antiviral vaccine or immunogeniccomposition and antiviral agents are administered simultaneously).Simultaneously includes administration contemporaneously or immediatelysequentially, that is during the same period of time. In certainembodiments, the one or more agents are administered simultaneously inthe same hour, or simultaneously in the same day. Sequential orsubstantially simultaneous administration of each therapeutic agent canbe effected by any appropriate route including, but not limited to, oralroutes, intravenous routes, sub-cutaneous routes, intramuscular routes,direct absorption through mucous membrane tissues (e.g., nasal, mouth,vaginal, and rectal), and ocular routes (e.g., intravitreal,intraocular, etc.). The therapeutic agents can be administered by thesame route or by different routes. For example, one component of aparticular combination may be administered by intravenous injectionwhile the other component(s) of the combination may be administeredintramuscularly only. The components may be administered in anytherapeutically effective sequence. A “combination” embraces groups ofcompounds or non-small chemical compound therapies useful as part of acombination therapy. In some embodiments, the therapeutic agent is ananti-retroviral therapy, (such as one or a combination of efavirenz,lamivudine and tenofovir disoproxil fumarate) or anti-flu therapy (suchas TamiFlu®). In some embodiments, the therapeutic agent is one or acombiantion of: abacavir/dolutegravir/lamivudine (Triumeq),dolutegravir/rilpivirine (Juluca),elvitegravir/cobicistat/emtricitabine/tenofovir disoproxil fumarate(Stribild), elvitegravir/cobicistat/emtricitabine/tenofovir alafenami de(Genvoya), efavirenz/emtricitabine/tenofovir disoproxil fumarate(Atripla), emtricitabine/rilpivirine/tenofovir disoproxil fumarate(Complera), emtricitabine/rilpivirine/tenofovir alafenamide (Odefsey),bictegravir, emtricitabine, and tenofovir alafenamide (Biktarvy). Insome embodiments, the therapeutic agent is one or a combination of areverse transcrioptase inhibitor of a retrovirus such as efavirenz(Sustiva), etravirine (Intelence), nevirapine (Viramune), nevirapineextended-release (Viramune XR), rilpivirine (Edurant), delavirdinemesylate (Rescriptor). In some embodiments, the therapeutic agent is oneor a combination of a protease inhibitor of a retrovirus, such as:atazanavir/cobicistat (Evotaz), darunavir/cobicistat (Prezcobix),lopinavir/ritonavir (Kaletra), ritonavir (Norvir), atazanavir (Reyataz),darunavir (Prezista), fosamprenavir (Lexiva), tipranavir (Aptivus).

As used herein, “expression” refers to the process by which apolynucleotide is transcribed from a DNA template (such as into and mRNAor other RNA transcript) and/or the process by which a transcribed mRNA(or administered mRNA) is translated into peptides, polypeptides, orproteins. Transcripts and encoded polypeptides may be collectivelyreferred to as “gene product.” If the polynucleotide is derived fromgenomic DNA, expression may include splicing of the mRNA in a eukaryoticcell. In some embodiments, the at least one expressible nucleic acidsequence comprises only DNA nucleotides, RNA nucleotides or comprisesboth RNA and DNA nucleotides. In some embodiments, the at least oneexpressible nucleic acid consist of RNA. In some embodiments, the atleast one expressible nucleic acid consist of DNA.

The terms “functional fragment” means any portion of a polypeptide ornucleic acid sequence from which the respective full-length polypeptideor nucleic acid relates that is of a sufficient length and has asufficient structure to confer a biological affect that is at leastsimilar or substantially similar to the full-length polypeptide ornucleic acid upon which the fragment is based. In some embodiments, afunctional fragment is a portion of a full-length or wild-type nucleicacid sequence that encodes any one of the nucleic acid sequencesdisclosed herein, and said portion encodes a polypeptide of a certainlength and/or structure that is less than full-length but encodes adomain that still biologically functional as compared to the full-lengthor wild-type protein. In some embodiments, the functional fragment mayhave a reduced biological activity, about equivalent biologicalactivity, or an enhanced biological activity as compared to thewild-type or full-length polypeptide sequence upon which the fragment isbased (such wild-type or full length sequences “reference sequences” oreach individually a “reference sequence”). In some embodiments, thefunctional fragment is derived from the sequence of an organism, such asa human. In such embodiments, the functional fragment may retain about99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% sequence identity tothe wild-type human sequence upon which the sequence is derived. In someembodiments, the functional fragment may retain about 85%, 80%, 75%,70%, 65%, or 60% sequence identity to the wild-type sequence upon whichthe sequence is derived.

By “fragment” is meant a portion of a polypeptide or nucleic acidmolecule. This portion contains, preferably, at least about 5%, 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, or about 90% of the entire length ofthe reference nucleic acid molecule or polypeptide. A fragment maycontain about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400,500, 600, 700, 800, 900, 1000 or more nucleotides or amino acids.

“Optional” or “optionally” means that the subsequently described event,circumstance, or material may or may not occur or be present, and thatthe description includes instances where the event, circumstance, ormaterial occurs or is present and instances where it does not occur oris not present.

The phrase “and/or,” as used herein in the specification and in theclaims, should be understood to mean “either or both” of the elements soconjoined, i.e., elements that are conjunctively present in some casesand disjunctively present in other cases. Other elements may optionallybe present other than the elements specifically identified by the“and/or” clause, whether related or unrelated to those elementsspecifically identified unless clearly indicated to the contrary. Thus,as a non-limiting example, a reference to “A and/or B,” when used inconjunction with open-ended language such as “comprising” can refer, insome embodiments, to A without B (optionally including elements otherthan B); in another embodiments, to B without A (optionally includingelements other than A); in yet another embodiments, to both A and B(optionally including other elements); etc.

As used herein in the specification and in the claims, “or” should heunderstood to have the same meaning as “and/or” as defined above. Forexample, when separating items in a list, “or” or “and/or” shall beinterpreted as being inclusive, i.e., the inclusion of at least one, butalso including more than one, of a number or list of elements, and,optionally, additional unlisted items. Only terms clearly indicated tothe contrary, such as “only one of” or “exactly one of,” or, when usedin the claims, “consisting of,” will refer to the inclusion of exactlyone element of a number or list of elements. In general, the term “or”as used herein shall only be interpreted as indicating exclusivealternatives (i.e. “one or the other but not both”) when preceded byterms of exclusivity, “either,” “one of,” “only one of,” or “exactly oneof” “Consisting essentially of,” when used in the claims, shall have itsordinary meaning as used in the field of patent law.

As used herein an “antigen” is meant to refer to any substance thatelicits an immune response.

As used herein, the term “electroporation,” “electro-permeabilization,”or “electro-kinetic enhancement” (“EP”), are used interchangeably andare meant to refer to the use of a transmembrane electric field pulse toinduce microscopic pathways (pores) in a bio-membrane; their presenceallows biomolecules such as plasmids, oligonucleotides, siRNA, drugs,ions, and/or water to pass from one side of the cellular membrane to theother. In some of the disclosed methods of treatment or prevention, themethod comprises a step of electroporation of a subject's tissue for asufficient time and with a sufficient electrical field capable ofinducing uptake of the pharmaceutical compositions disclosed herein intothe antigen-presenting cells. In some embodiments, the cells are antigenpresenting cells.

The term “pharmaceutically acceptable excipient,” “pharmaceuticallyacceptable carrier” or “pharmaceutically acceptable diluent” as usedherein is meant to refer to an excipient, carrier or diluent that can beadministered to a subject, together with an agent or the pharmaceuticalcompositions disclosed herein, and which is inert or fails to eliminatethe pharmacological activity of the active agent of the pharmaceuticalcomposition. In some embodiments, the pharmaceutically acceptablecarrier does fails to destroy or is incapable of eliminating thepharmacological activity of an active agent/vaccine and and is nontoxicwhen administered in doses sufficient to deliver a therapeutic amount ofthe active agent. The term “pharmaceutically acceptable salt” of nucleicacids as used herein may be an acid or base salt that is generallyconsidered in the art to be suitable for use in contact with the tissuesof human beings or animals without excessive toxicity, irritation,allergic response, or other problem or complication. Such salts includemineral and organic acid salts of basic residues such as amines, as wellas alkali or organic salts of acidic residues such as carboxylic acids.Specific pharmaceutical salts include, but are not limited to, salts ofacids such as hydrochloric, phosphoric, hydrobromic, malic, glycolic,fumaric, sulfuric, sulfamic, suifanilic, formic, toluenesulfonie,methanesulfonic, benzene sulfonic, ethane disulfonic, 2-hydroxyethylsulfonic, nitric, benzoic, 2-acetoxybenzoic, citric, tartaric, lactic,stearic, salicylic, glutamic, ascorbic, pamoic, succinic, fumaric,maleic, propionic, hydroxymaleic, hydroiodic, phenyiacetic, alkanoicsuch as acetic, HOOC—(CH₂)n-COOH where n is 0-4, and the like.Similarly, pharmaceutically acceptable cations include, but are notlimited to sodium, potassium, calcium, aluminum, lithium and ammonium.Those of ordinary skill in the art will recognize from this disclosureand the knowledge in the art that further pharmaceutically acceptablesalts for the pooled viral specific antigens or polynucleotides providedherein, including those listed by Remington's Pharmaceutical Sciences,17th ed., Mack Publishing Company, Easton, Pa., p. 1418 (1985). Ingeneral, a pharmaceutically acceptable acid or base salt can besynthesized from a parent compound that contains a basic or acidicmoiety by any conventional chemical method. Briefly, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in anappropriate solvent.

As used herein, the terms “prevent,” “preventing,” “prevention,”“prophylactic treatment,” and the like, are meant to refer to reducingthe probability of developing a disease or condition in a subject, whodoes not have, but is at risk of or susceptible to developing a diseaseor condition.

As used herein, the term “purified” means that the polynucleotide orpolypeptide or fragment, variant, or derivative thereof is substantiallyfree of other biological material with which it is naturally associated,or free from other biological materials derived, e.g., from arecombinant host cell that has been genetically engineered to expressthe polypeptide of the present disclosure. That is, e.g., a purifiedpolypeptide of the present disclosure is a polypeptide that is at leastfrom about 70 to 100% pure, i.e., the polypeptide is present in acomposition wherein the polypeptide constitutes from about 70 to about100% by weight of the total composition. In some embodiments, thepurified polypeptide of the present disclosure is from about 75% toabout 99% by weight pure, from about 80% to about 99% by weight pure,from about 90 to about 99% by weight pure, or from about 95% to about99% by weight pure.

The terms “subject,” “individual,” and “patient” are usedinterchangeably herein to refer to a vertebrate, preferably a mammal,more preferably a human. Mammals include, but are not limited to,murine, simians, humans, farm animals, cows, pigs, goats, sheep, horses,dogs, sport animals, and pets. Tissues, cells and their progeny obtainedin vivo or cultured in vitro are also encompassed by the definition ofthe term “subject.” The term “subject” is also used throughout thespecification in some embodiments to describe an animal from which acell sample is taken or an animal to which a disclosed cell or nucleicacid sequences have been administered. In some embodiment, the subjectis a human. For treatment of those conditions which are specific for aspecific subject, such as a human being, the term “patient” may beinterchangeably used. In some instances in the description of thepresent disclosure, the term “patient” will refer to human patientssuffering from a particular disease or disorder. In some embodiments,the subject may be a non-human animal. The term “mammal” encompassesboth humans and non-humans and includes but is not limited to humans,non-human primates, canines, felines, murine, bovines, equines, caprine,and porcines.

The term “therapeutic effect” as used herein is meant to refer to someextent of relief of one or more of the symptoms of a disorder (e.g.,SARS-CoV-2 infection) or its associated pathology. A “therapeuticallyeffective amount” as used herein is meant to refer to an amount of anagent which is effective, upon single or multiple dose administration(such as a first, second and/or third booster) to the cell or subject,in prolonging the survivability of the patient with such a disorder,reducing one or more signs or symptoms of the disorder, preventing ordelaying, and the like beyond that expected in the absence of suchtreatment. A “therapeutically effective amount” is intended to qualifythe amount required to achieve a therapeutic effect. A physician orveterinarian having ordinary skill in the art can readily determine andprescribe the “therapeutically effective amount” (e.g., ED50) of thepharmaceutical composition required. For example, the physician orveterinarian could start doses of the compounds of the presentdisclosure employed in a pharmaceutical composition at levels lower thanthat required in order to achieve the desired therapeutic effect andgradually increase the dosage until the desired effect is achieved.

The terms “treat,” “treated,” “treating,” “treatment,” and the like asused herein are meant to refer to reducing or ameliorating a disorderand/or symptoms associated therewith (e.g., a viral infection).“Treating” can refer to administration of the DNA and/or RNA vaccinesdescribed herein to a subject after the onset, or suspected onset, of aviral infection. “Treating” includes the concepts of “alleviating,”which refers to lessening the frequency of occurrence or recurrence, orthe severity, of any symptoms or other ill effects related to a virusand/or the side effects associated with viral therapy. The term“treating” also encompasses the concept of “managing” which refers toreducing the severity of a particular disease or disorder in a patientor delaying its recurrence, e.g., lengthening the period of remission ina patient who had suffered from the disease. It is appreciated that,although not precluded, treating a disorder or condition does notrequire that the disorder, condition, or symptoms associated therewithbe completely eliminated.

For any therapeutic agent described herein the therapeutically effectiveamount may be initially determined from preliminary in vitro studiesand/or animal models. A therapeutically effective dose may also bedetermined from human data. The applied dose can be adjusted based onthe relative bioavailability and potency of the administered agent.Adjusting the dose to achieve maximal efficacy based on the methodsdescribed above and other well-known methods is within the capabilitiesof the ordinarily skilled artisan. General principles for determiningtherapeutic effectiveness, which may be found in Chapter 1 of Goodmanand Gilman's The Pharmacological Basis of Therapeutics, 10th Edition,McGraw-Hill (New York) (2001), incorporated herein by reference, aresummarized below. Pharmacokinetic principles provide a basis formodifying a dosage regimen to obtain a desired degree of therapeuticefficacy with a minimum of unacceptable adverse effects. In situationswhere the drug's plasma concentration can be measured and related to thetherapeutic window, additional guidance for dosage modification can beobtained. Drug products are considered to be pharmaceutical equivalentsif they contain the same active ingredients and are identical instrength or concentration, dosage form, and route of administration. Twopharmaceutically equivalent drug products are considered to bebioequivalent when the rates and extents of bioavailability of theactive ingredient in the two products are not significantly differentunder suitable test conditions.

The terms “polynucleotide,” “oligonucleotide” and “nucleic acid” areused interchangeably throughout and include DNA molecules (e.g., cDNA orgenomic DNA), RNA molecules (e.g., mRNA), analogs of the DNA or RNAgenerated using nucleotide analogs (e.g., peptide nucleic acids andnon-naturally occurring nucleotide analogs), and hybrids thereof. Thenucleic acid molecule can be single-stranded or double-stranded. In someembodiments, the nucleic acid molecules of the disclosure comprise acontiguous open reading frame encoding an antibody, or a fragmentthereof, as described herein. “Nucleic acid” or “oligonucleotide” or“polynucleotide” as used herein may mean at least two nucleotidescovalently linked together. The depiction of a single strand alsodefines the sequence of the complementary strand. Thus, a nucleic acidalso encompasses the complementary strand of a depicted single strand.Many variants of a nucleic acid may be used for the same purpose as agiven nucleic acid. Thus, a nucleic acid also encompasses substantiallyidentical nucleic acids and complements thereof. A single strandprovides a probe that may hybridize to a target sequence under stringenthybridization conditions. Thus, a nucleic acid also encompasses a probethat hybridizes under stringent hybridization conditions. Nucleic acidsmay be single stranded or double stranded, or may contain portions ofboth double stranded and single stranded sequence. The nucleic acid maybe DNA, both genomic and cDNA, RNA, or a hybrid, where the nucleic acidmay contain combinations of deoxyribo- and ribo-nucleotides, andcombinations of bases including uracil, adenine, thymine, cytosine,guanine, inosine, xanthine hypoxanthine, isocytosine and isoguanine.Nucleic acids may be obtained by chemical synthesis methods or byrecombinant methods. A nucleic acid will generally containphosphodiester bonds, although nucleic acid analogs maybe included thatmay have at least one different linkage, e.g., phosphoramidate,phosphorothioate, phosphorodithioate, or 0-methylphosphoroamiditelinkages and peptide nucleic acid backbones and linkages. Other analognucleic acids include those with positive backbones; non-ionicbackbones, and non-ribose backbones, including those described in U.S.Pat. Nos. 5,235,033 and 5,034,506, which are incorporated by referencein their entireties.

Nucleic acids containing one or more non-naturally occurring or modifiednucleotides are also included within one definition of nucleic acids.The modified nucleotide analog may he located for example at the 5′-endand/or the 3′-end of the nucleic acid molecule. Representative examplesof nucleotide analogs may be selected from sugar- or backbone-modifiedribonucleotides. It should be noted, however, that alsonucleobase-modified ribonucleotides, i.e. ribonucleotides, containing anon-naturally occurring nucleobase instead of a naturally occurringnucleobase such as uridines or cytidines modified at the 5-position,e.g. 5-(2-amino)propyl uridine, 5-bromo uridine; adenosines andguanosines modified at the 8-position, e.g. 8-bromo guanosine; deazanucleotides, e.g. 7-deaza-adenosine; 0- and N-alkylated nucleotides,e.g. N6-methyl adenosine are suitable. The 2′-OH-group may be replacedby a group selected from H, OR, R, halo, SH, SR, NH₂, NHR, N₂ or CN,wherein R is C₁-C₆ alkyl, alkenyl or alkynyl and halo is F, Cl, Br or I.Modified nucleotides also include nucleotides conjugated withcholesterol through, e.g., a hydroxyprolinol linkage as described inKrutzfeldt et al., Nature (Oct. 30, 2005), Soutschek et al., Nature432:173-178 (2004), and U.S. Patent Publication No. 20050107325, whichare incorporated herein by reference in their entireties. Modifiednucleotides and nucleic acids may also include locked nucleic acids(LNA), as described in U.S. Pat. No. 20020115080, which is incorporatedherein by reference. Additional modified nucleotides and nucleic acidsare described in U.S. Patent Publication No. 20050182005, which isincorporated herein by reference in its entirety. Modifications of theribose-phosphate backbone may be done for a variety of reasons, e.g., toincrease the stability and half-life of such molecules in physiologicalenvironments, to enhance diffusion across cell membranes, or as probeson a biochip. Mixtures of naturally occurring nucleic acids and analogsmay be made; alternatively, mixtures of different nucleic acid analogs,and mixtures of naturally occurring nucleic acids and analogs may bemade. In some embodiments, the expressible nucleic acid sequence is inthe form of DNA. In some embodiments, the expressible nucleic acid is inthe form of RNA with a sequence that encodes the polypeptide sequencesdisclosed herein and, in some embodiments, the expressible nucleic acidsequence is an RNA/DNA hybrid molecule that encodes any one or pluralityof polypeptide sequences disclosed herein.

As used herein, the term “nucleic acid molecule” is a molecule thatcomprises one or more nucleotide sequences that encode one or moreproteins. In some embodiments, a nucleic acid molecule comprisesinitiation and termination signals operably linked to regulatoryelements including a promoter and polyadenylation signal capable ofdirecting expression in the cells of the individual to whom the nucleicacid molecule is administered. In some embodiments, the nucleic acidmolecule also includes a plasmid containing one or more nucleotidesequences that encode one or a plurality of viral antigens. In someembodiments, the disclosure relates to a pharmaceutical compositioncomprising a first, second, third or more nucleic acid molecule, each ofwhich encoding one or a plurality of viral antigens and at least one ofeach plasmid comprising one or more of the compositions disclosedherein.

The terms “polypeptide,” “peptide” and “protein” are usedinterchangeably herein to refer to polymers of amino acids of anylength. The polymer may be linear or branched, it may comprise modifiedamino acids, and it may be interrupted by non-natural amino acids orchemical groups that are not amino acids. The terms also encompass anamino acid polymer that has been modified; for example, disulfide bondformation, glycosylation, lipidation, acetylation, phosphorylation, orany other manipulation, such as conjugation with a labeling component.As used herein the term “amino acid” includes natural and/or unnaturalor synthetic amino acids, including glycine and both the D or L opticalisomers, and amino acid analogs and peptidomimetics.

The “percent identity” or “percent homology” of two polynucleotide ortwo polypeptide sequences is determined by comparing the sequences usingthe GAP computer program (a part of the GCG Wisconsin Package, version10.3 (Accelrys, San Diego, Calif.)) using its default parameters.“Identical” or “identity” as used herein in the context of two or morenucleic acids or amino acid sequences, may mean that the sequences havea specified percentage of residues that are the same over a specifiedregion. The percentage may be calculated by optimally aligning the twosequences, comparing the two sequences over the specified region,determining the number of positions at which the identical residueoccurs in both sequences to yield the number of matched positions,dividing the number of matched positions by the total number ofpositions in the specified region, and multiplying the result by 100 toyield the percentage of sequence identity. In cases where the twosequences are of different lengths or the alignment produces one or morestaggered ends and the specified region of comparison includes only asingle sequence, the residues of single sequence are included in thedenominator but not the numerator of the calculation. When comparing DNAand RNA, thymine (T) and uracil (U) may be considered equivalent.Identity may he performed manually or by using a computer sequencealgorithm such as BLAST or BLAST 2.0. Briefly, the BLAST algorithm,which stands for Basic Local Alignment Search Tool is suitable fordetermining sequence similarity. Software for performing BLAST analysesis publicly available through the National Center for BiotechnologyInformation (ncbi.nlm.nih.gov). This algorithm involves firstidentifying high scoring sequence pair (HSPs) by identifying short wordsof length Win the query sequence that either match or satisfy somepositive-valued threshold score T when aligned with a word of the samelength in a database sequence. T is referred to as the neighborhood wordscore threshold (Altschul et al., supra). These initial neighborhoodword hits act as seeds for initiating searches to find HSPs containingthem. The word hits are extended in both directions along each sequencefor as far as the cumulative alignment score can be increased. Extensionfor the word hits in each direction are halted when: 1) the cumulativealignment score falls off by the quantity X from its maximum achievedvalue; 2) the cumulative score goes to zero or below, due to theaccumulation of one or more negative-scoring residue alignments; or 3)the end of either sequence is reached. The Blast algorithm parameters W,T and X determine the sensitivity and speed of the alignment. The Blastprogram uses as defaults a word length (W) of 11, the BLOSUM62 scoringmatrix (see Henikoff et al., Proc. Natl. Acad. Sci. USA, 1992, 89,10915-10919, which is incorporated herein by reference in its entirety)alignments (B) of 50, expectation (E) of 10, M=5, N=4, and a comparisonof both strands. The BLAST algorithm (Karlin et al., Proc. Natl. Acad.Sci. USA, 1993, 90, 5873-5787, which is incorporated herein by referencein its entirety) and Gapped BLAST perform a statistical analysis of thesimilarity between two sequences. One measure of similarity provided bythe BLAST algorithm is the smallest sum probability (P(N)), whichprovides an indication of the probability by which a match between twonucleotide sequences would occur by chance. For example, a nucleic acidis considered similar to another if the smallest sum probability incomparison of the test nucleic acid to the other nucleic acid is lessthan about 1, less than about 0.1, less than about 0.01, and less thanabout 0.001. Two single-stranded polynucleotides are “the complement” ofeach other if their sequences can be aligned in an anti-parallelorientation such that every nucleotide in one polynucleotide is oppositeits complementary nucleotide in the other polynucleotide, without theintroduction of gaps, and without unpaired nucleotides at the 5′ or the3′ end of either sequence. A polynucleotide is “complementary” toanother polynucleotide if the two polynucleotides can hybridize to oneanother under moderately stringent conditions. Thus, a polynucleotidecan be complementary to another polynucleotide without being itscomplement.

The term “hybridization” or “hybridizes” as used herein refers to theformation of a duplex between nucleotide sequences that are sufficientlycomplementary to form duplexes via Watson-Crick base pairing. Twonucleotide sequences are “complementary” to one another when thosemolecules share base pair organization homology. “Complementary”nucleotide sequences will combine with specificity to form a stableduplex under appropriate hybridization conditions. For instance, twosequences are complementary when a section of a first sequence can bindto a section of a second sequence in an anti-parallel sense wherein the3′-end of each sequence binds to the 5′-end of the other sequence andeach A, T(U), G and C of one sequence is then aligned with a T(U), A, Cand G, respectively, of the other sequence. RNA sequences can alsoinclude complementary G=U or U=G base pairs. Thus, two sequences neednot have perfect homology to be “complementary.” Usually two sequencesare sufficiently complementary when at least about 90% (preferably atleast about 95%) of the nucleotides share base pair organization over adefined length of the molecule.

By “substantially identical” is meant nucleic acid molecule (orpolypeptide) exhibiting at least about 50% identity to a reference aminoacid sequence (for example, any one of the amino acid sequencesdescribed herein) or nucleic acid sequence (for example, any one of thenucleic acid sequences described herein). In some embodiments, such asequence is at least about 60%, 70%, 80% or 85%, 90%, 95% or even 99%identical at the amino acid level or nucleic acid to the sequence usedfor comparison.

A nucleotide sequence is “operably linked” to a regulatory sequence ifthe regulatory sequence affects the expression (e.g., the level, timing,or location of expression) of the nucleotide sequence. A “regulatorysequence” is a nucleic acid that affects the expression (e.g., thelevel, timing, or location of expression) of a nucleic acid to which itis operably linked. The regulatory sequence can, for example, exert itseffects directly on the regulated nucleic acid, or through the action ofone or more other molecules (e.g., polypeptides that bind to theregulatory sequence and/or the nucleic acid). Examples of regulatorysequences include promoters, enhancers and other expression controlelements (e.g., polyadenylation signals). Further examples of regulatorysequences are described in, for example, Goeddel, 1990, Gene ExpressionTechnology: Methods in Enzymology 185, Academic Press, San Diego, Calif.and Baron et al., 1995, Nucleic Acids Res. 23:3605-06.

A “vector” is a nucleic acid that can be used to introduce anothernucleic acid linked to it into a cell. One type of vector is a“plasmid,” which refers to a linear or circular double stranded DNAmolecule into which additional nucleic acid segments can be ligated.Another type of vector is a viral vector (e.g., replication defectiveretroviruses, adenoviruses and adeno-associated viruses), comprisingadditional, exogenous DNA, RNA or hybrid DNA or RNA molecules that canbe introduced into the viral genome. Certain vectors are capable ofautonomous replication in a host cell into which they are introduced(e.g., bacterial vectors comprising a bacterial origin of replicationand episomal mammalian vectors). Other vectors (e.g., non-episomalmammalian vectors) are integrated into the genome of a host cell uponintroduction into the host cell, and thereby are replicated along withthe host genome. An “expression vector” is a type of vector that candirect the expression of a chosen polynucleotide. The disclosure relatesto any one or plurality of vectors that comprise nucleic acid sequencesencoding any one or plurality of amino acid sequence disclosed herein.In some embodiments, the expression vector includes from about 30 toabout 100,000 nucleotides (e.g., from about 30 to about 50, from about30 to about 100, from about 30 to about 250, from about 30 to about 500,from about 30 to about 1,000, from about 30 to about 1,500, from about30 to about 3,000, from about 30 to about 5,000, from about 30 to about7,000, from about 30 to about 10,000, from about 30 to about 25,000,from about 30 to about 50,000, from about 30 to about 70,000, from about100 to about 250, from about 100 to about 500, from about 100 to about1,000, from about 100 to about 1,500, from about 100 to about 3,000,from about 100 to about 5,000, from about 100 to about 7,000, from about100 to about 10,000, from about 100 to about 25,000, from about 100 toabout 50,000, from about 100 to about 70,000, from about 100 to about100,000, from about 500 to about 1,000, from about 500 to about 1,500,from about 500 to about 2,000, from about 500 to about 3,000, from about500 to about 5,000, from about 500 to about 7,000, from about 500 toabout 10,000, from about 500 to about 25,000, from about 500 to about50,000, from about 500 to about 70,000, from about 500 to about 100,000,from about 1,000 to about 1,500, from about 1,000 to about 2,000, fromabout 1,000 to about 3,000, from about 1,000 to about 5,000, from about1,000 to about 7,000, from about 1,000 to about 10,000, from about 1,000to about 25,000, from about 1,000 to about 50,000, from about 1,000 toabout 70,000, from about 1,000 to about 100,000, from about 1,500 toabout 3,000, from about 1,500 to about 5,000, from about 1,500 to about7,000, from about 1,500 to about 10,000, from about 1,500 to about25,000, from about 1,500 to about 50,000, from about 1,500 to about70,000, from about 1,500 to about 100,000, from about 2,000 to about3,000, from about 2,000 to about 5,000, from about 2,000 to about 7,000,from about 2,000 to about 10,000, from about 2,000 to about 25,000, fromabout 2,000 to about 50,000, from about 2,000 to about 70,000, and fromabout 2,000 to about 100,000 nucleotides).

The term “vaccine” as used herein is meant to refer to a compositioncapable of generating immunity for the prophylaxis and/or treatment ofdiseases (e.g., viral infections). In some embodiments, the vaccine is acomposition capable of generating therapeutically effective immunity forthe prophylaxis or treatment of Coronaviridae infection or propagationin a subject. Accordingly, vaccines are medicaments which compriseantigens in protein and/or nucleic acid forms and are in animals forgenerating specific defense and protective substance by vaccination. A“vaccine composition” can include a pharmaceutically acceptableexcipient, carrier or diluent. A “vaccine composition” or “nucleic acidvaccine composition” as used herein can comprise a DNA vaccine, a RNAvaccine or a combination thereof.

“Variants” are intended to mean substantially similar sequences. Fornucleic acid molecules, a variant comprises a nucleic acid moleculehaving deletions (i.e., truncations) at the 5′ and/or 3′ end; deletionand/or addition of one or more nucleotides at one or more internal sitesin the native polynucleotide; and/or substitution of one or morenucleotides at one or more sites in the native polynucleotide. As usedherein, a “native” nucleic acid molecule or polypeptide comprises anaturally occurring or endogenous nucleotide sequence or amino acidsequence, respectively. For nucleic acid molecules, conservativevariants include those sequences that, because of the degeneracy of thegenetic code, encode the amino acid sequence of one of the polypeptidesof the disclosure. Variant nucleic acid molecules also includesynthetically derived nucleic acid molecules, such as those generated,for example, by using site-directed mutagenesis but which still encode aprotein of the disclosure. Generally, variants of a particular nucleicacid molecule of the disclosure will have at least about 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequenceidentity to that particular polynucleotide as determined by sequencealignment programs and parameters as described elsewhere herein.Variants of a particular nucleic acid molecule of the disclosure (i.e.,the reference DNA sequence) can also be evaluated by comparison of thepercent sequence identity between the polypeptide encoded by a variantnucleic acid molecule and the polypeptide encoded by the referencenucleic acid molecule. Percent sequence identity between any twopolypeptides can be calculated using sequence alignment programs andparameters described elsewhere herein. Where any given pair of nucleicacid molecule of the disclosure is evaluated by comparison of thepercent sequence identity shared by the two polypeptides that theyencode, the percent sequence identity between the two encodedpolypeptides is at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or more sequence identity. In someembodiments, the term “variant” protein is intended to mean a proteinderived from the native protein by deletion (so-called truncation) ofone or more amino acids at the N-terminal and/or C-terminal end of thenative protein; deletion and/or addition of one or more amino acids atone or more internal sites in the native protein; or substitution of oneor more amino acids at one or more sites in the native or wild-typeprotein upon which the variant structure is based. Variant proteinsencompassed by the present disclosure are biologically active, that isthey continue to possess the desired biological activity of the nativeprotein as described herein. Such variants may result from, for example,genetic polymorphism or from human manipulation. Biologically activevariants of a protein of the disclosure will have at least about 70%,75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or moresequence identity to the amino acid sequence for the native protein asdetermined by sequence alignment programs and parameters describedelsewhere herein. A biologically active variant of a protein of thedisclosure may differ from that protein by as few as 1-15 amino acidresidues, as few as 1-10, such as 6-10, as few as 5, as few as 4, 3, 2,or even 1 amino acid residue. The proteins or polypeptides of thedisclosure may be altered in various ways including amino acidsubstitutions, deletions, truncations, and insertions. Methods for suchmanipulations are generally known in the art. For example, amino acidsequence variants and fragments of the proteins can be prepared bymutations in the nucleic acid sequence that encode the amino acidsequence recombinantly. In some embodiments, the nucleic acid moleculesor the nucleic acid sequences comprise conservative mutations of 1, 2,3, 4, 5, 6, 7, 8, 9, 10 or more nucleotides.

As used herein, “SARS-CoV-2” refers to severe acute respiratorysyndrome-related coronavirus-2 or a subtype or variant thereof.Non-limiting examples of the SARS-CoV-2 virus, variants and subtypesthereof are described, for example, in Morais et al. (Sci. Rep., 2020,10(1):18289), Zhao et al. (PLoS Comput Biol., 2020, 16(9):e1008269),Forster et al. (PNAS, 2020, 117(17):9241-9243), and Tang et al.(National Science Review, 2020, 7(6):1012-1023), all of which areincorporated by reference herein. In some embodiments, the SARS-CoV-2 isone of the 6 subtypes and 10 tentative subtypes identified in Morais etal. (Sci. Rep., 2020, 10(1):18289). In some embodiments, the SARS-CoV-2is one of the 3 SARS-CoV-2 variants identified in Foster et al. (PNAS,2020, 117(17):9241-9243). In some embodiments, the SARS-CoV-2 is one ofthe two SARS-CoV-2 subtypes identified in Tang et al. (National ScienceReview, 2020, 7(6):1012-1023). In some embodiments, the SARS-CoV-2 isSARS-CoV-2 hCoV-19/Australia/VIC01/2020 or a variant thereof. In someembodiments, SARS-COV-2 comprises the sequences as described in NCBIReference Sequence: NC 045512.2 or a variant thereof. In someembodiments, SARS-CoV-2 comprises the sequence as described in GenBank:MN908947.3 or a variant thereof. In some embodiments, the SARS-Cov-2variant is the B.1.1.7 varian, also referred to as lineage B.1.1.7, VOC202012/01 or 20I/501Y.V1. In some embodiments, the SARS-Cov-2 variant isthe B.1.351 variant, also referred to as B.1.351 lineage. In someembodiments, the SARS-Cov-2 variant is the B.1.1.28 subclade (renamed“P.1”). In some embodiments, the SARS-Cov-2 variant is the B.1.1.7variant, also referred to as B.1.1.7 lineage or 20I/501Y.V1.

As used herein, a “scaffold domain” refers to a region of a polypeptidethat structurally facilitates, or acts as a scaffold for, presentationof one or a plurality of viral antigens. In some embodiments, a scaffolddomain of the disclosure comprises a self-assembling polypeptide which,when expressed, forms an inner scaffold or core upon which one or aplurality of viral antigens are displayed. In some embodiments, theself-assemblying peptide forms a three dimensional shape with identicalsingle polypeptide surfaces. In some embodiments, the scaffold domanforms a secondary and/or tertiary structure that is a polyhedron. Insome embodiments, the scaffold domain encodes a monomer or series ofmonomers that form a 7, 14, 20, 28 or 60 faces of a polyhedron uponwhich antigens are encoded or displayed.

As used herein, the term “trimer” refers to a protein complex formed bythree individual monomers. In some embodiments, the monomer subunits ofthe trimer are bound together to form a trimer by non-covalentprotein-protein interactions.

Finally, it should be understood that all of the individual values andsub-ranges of values contained within an explicitly disclosed range arealso specifically contemplated and should be considered disclosed unlessthe context specifically indicates otherwise. The foregoing appliesregardless of whether in particular cases some or all of theseembodiments are explicitly disclosed.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meanings as commonly understood by one of skill in the artto which the disclosed method and compositions belong. Although anymethods and materials similar or equivalent to those described hereincan be used in the practice or testing of the present method andcompositions, the particularly useful methods, devices, and materialsare as described. Publications cited herein and the material for whichthey are cited are hereby specifically incorporated by reference.Nothing herein is to be construed as an admission that the presentdisclosure is not entitled to antedate such disclosure by virtue ofprior disclosure. No admission is made that any reference constitutesprior art. The discussion of references states what their authorsassert, and applicants reserve the right to challenge the accuracy andpertinency of the cited documents. It will be clearly understood that,although a number of publications are referred to herein, such referencedoes not constitute an admission that any of these documents forms partof the common general knowledge in the art.

Throughout the description and claims of this specification, the word“comprise” and variations of the word, such as “comprising” and“comprises,” means “including but not limited to,” and is not intendedto exclude, for example, other additives, components, integers or steps.In particular, in methods stated as comprising one or more steps oroperations it is specifically contemplated that each step comprises whatis listed (unless that step includes a limiting term such as “consistingof”), meaning that each step is not intended to exclude, for example,other additives, components, integers or steps that are not listed inthe step.

A. Nucleic Acid Compositions

Disclosed are compositions and pharmaceutical compositions comprisingone or plurality of expressible nucleic acid sequences. In someembodiments, the expressible nucleic acid sequence is a DNA. In otherembodiments, the expressible nucleic acid sequence is a RNA. In someembodiments, the expressible nucleic acid is operably linked to one or aplurality of regulatory sequences. In some embodiments, the expressiblenucleic acid sequence is comprised and forms a part of a nucleic acidmolecule, such as a vector or plasmid.

In one aspect, the expressible nucleic acid sequence of the disclosurecomprises a first nucleic acid sequence encoding a scaffold domaincomprising a self-assembling polypeptide or a pharmaceuticallyacceptable salt thereof, and a second nucleic acid sequence encoding anantigen domain comprising a viral antigen or a pharmaceuticallyacceptable salt thereof. The self-assembling polypeptide is aself-assembling peptide that is expressed to facilitate presentation ofthe viral antigen. Transformed or transfected cells exposed to suchexpressible nucleic acid sequences can produce the self-assemblingpeptide which is enveloped by the viral antigens, thereby stimulatingthe viral antigen-specific immune response against the antigen. In someembodiments, the antigen-specific immune response is a therapeuticallyeffective immune response against the virus from which the antigen aminoacid sequence is obtained. In some embodiments, the viral antigenencoded by the expressible nucleic acid of the disclosure comprises acoronaviral antigen. In some embodiments, the expressible nucleic acidsequence further comprises a third nucleic acid sequence encoding aleader sequence or a pharmaceutically acceptable salt thereof. In someembodiments, the leader sequence is an IgE or IgG leader sequence. Insome embodiments, the expressible nucleic acid sequence furthercomprises a fourth nucleic acid sequence encoding a linker peptide or apharmaceutically acceptable salt thereof, wherein the fourth nucleicacid sequence is positioned between the first nucleic acid sequence andthe second nucleic acid sequence in the 5′ to 3′ orientation. In someembodiments, the expressible nucleic acid sequence comprises a firstnucleic acid sequence encoding a scaffold domain comprising aself-assembling polypeptide or a pharmaceutically acceptable saltthereof, a second nucleic acid sequence encoding an antigen domaincomprising a viral antigen or a pharmaceutically acceptable saltthereof, and a third nucleic acid sequence encoding a leader sequence ora pharmaceutically acceptable salt thereof. In some embodiments, theexpressible nucleic acid sequence comprises a first nucleic acidsequence encoding a scaffold domain comprising a self-assemblingpolypeptide or a pharmaceutically acceptable salt thereof, a secondnucleic acid sequence encoding an antigen domain comprising a viralantigen or a pharmaceutically acceptable salt thereof, a third nucleicacid sequence encoding a leader sequence or a pharmaceuticallyacceptable salt thereof, and a fourth nucleic acid sequence encoding alinker peptide or a pharmaceutically acceptable salt thereof, whereinthe fourth nucleic acid sequence is positioned between the first nucleicacid sequence and the second nucleic acid sequence in the 5′ to 3′orientation.

In some embodiments, the expressible nucleic acid sequence of thedisclosure comprises a nucleic acid sequence encoding a viral trimerpolypeptide, a functional fragment thereof or a pharmaceuticallyacceptable salt thereof. In some embodiments, the expressible nucleicacid sequence comprises, in a 5′ to 3′ orientation, a first nucleic acidsequence encoding a leader sequence or a pharmaceutically acceptablesalt thereof, and a second nucleic acid sequence encoding a viral trimerpolypeptide, a functional fragment or variant thereof or apharmaceutically acceptable salt thereof. In some embodiments, theleader sequence is an IgE or IgG leader sequence. In some embodiments,the expressible nucleic acid sequence comprises, in a 5′ to 3′orientation, a first nucleic acid sequence encoding a leader sequence ora pharmaceutically acceptable salt thereof, and a second nucleic acidsequence encoding a viral polypeptide that is a component of a viraltrimer, a functional fragment or variant thereof or a pharmaceuticallyacceptable salt thereof. In some embodiments, the viral polypeptide thatis a component of a viral trimer is a monomer of a viral trimer, suchthat, upon expression, the monomers spontaneously aggregate to form atrimeric viral polypeptide. In some embodiments, the viral trimerencoded by the expressible nucleic acid of the disclosure comprises acoronaviral trimer. In some embodiments, the viral trimer comprises thespike protein of SARS-CoV-2, a functional fragment or variant thereof ora pharmaceutically acceptable salt thereof.

In some embodiments, the nucleic acid sequences encoding the viralantigens or viral trimers comprised in the expressible nucleic acid ofthe disclosure comprise one or a plurality of mutations so to tailor thevaccine induced responses. Such mutations result in creating glycansites in the encoded polypeptide so that glycosylation events can beobtained. In some embodiments, such glycan modifications or mutationsdecrease the bottom reactivity. In some embodiments, such glycanmodifications or mutations increase antigen activity.

1. Leader Sequence

The expressible nucleic acid sequence of the present disclosureoptionally comprises a nucleic acid sequence encoding a leader sequence,a functional fragment or variant thereof or a pharmaceuticallyacceptable salt thereof. A “leader sequence” may from time to time referto a “signal peptide” and thus, the terms “leader sequence” and “signalpeptide” are used interchangeably herein and refer to an amino acidsequence that can be linked at the amino terminus of a protein set forthherein. Signal peptides/leader sequences typically direct localizationof a protein. Signal peptides/leader sequences used herein preferablyfacilitate secretion of the protein from the cell in which it isproduced. Signal peptides/leader sequences are often cleaved from theremainder of the protein, often referred to as the mature protein, uponsecretion from the cell. Signal peptides/leader sequences, when present,are linked at the N terminus of the protein. The presence of a leadersequence may be required for proper secretion of the viral antigen ortrimer encoded by the expressible nucleic acid sequence of thedisclosure.

A non-limiting example of the leader sequence is the IgE leader sequencecomprising the amino acid sequence of MDWTWILFLVAAATRVHS (SEQ ID NO: 1;also named “MD39”) encoded by one of the following nucleic acidsequences:

(SEQ ID NO: 2; “MD39”)atggactggacatggattctgttcctggtcgctgccgctacaagagtgca ttcc;(SEQ ID NO: 3; “CPG9.2”)atggattggacttggattctgttcctggtcgcagcagccacacgagtgca tagc; and(SEQ ID NO: 4) atggactggacctggattctgttcctggtggccgccgccacaagggtgca cagc.

Another non-limiting example of the leader sequence is the amino acidsequence of MDWTWRILFLVAAATGTHA (SEQ ID NO: 5) encoded by the nucleicacid sequence ofatggactggacctggagaatcctgttcctggtggccgccgccaccggcacacacgccgatacacacttccccatctgcatcttttgctgtggctgttgccataggtccaagtgtgggatgtgctgcaaaact (SEQ ID NO: 6).

Thus, in some embodiments when the leader sequence is present, theleader sequence comprises at least about 60%, 65%, 70%, 75%, 80%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%sequence identity to SEQ ID NO: 1 or SEQ ID NO: 5, or a functionalfragment or variant thereof. In some embodiments, the leader sequencecomprises the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 5, or afunctional fragment or variant thereof. In some embodiments, the leadersequence is encoded by a nucleic acid sequence comprising at least about60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 2, SEQ IDNO: 3, SEQ ID NO: 4 or SEQ ID NO: 6, or a functional fragment or variantthereof. In some embodiments, the leader sequence is encoded by thenucleic acid sequence of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 or SEQID NO: 6, or a functional fragment or variant thereof. In otherembodiments, the leader sequence is encoded by a nucleic acid sequencecomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequenceidentity to a nucleic acid sequence that is complementary to SEQ ID NO:2, SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 6, or a functional fragmentor variant thereof. In some embodiments, the leader sequence is encodedby a nucleic acid sequence that is complementary to the nucleic acidsequence of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 6, ora functional fragment or variant thereof

2. Self-Assembling Polypeptide

The disclosure relates to an expressible nucleic acid sequencecomprising at least one nucleic acid sequence encoding a scaffold domaincomprising a self-assembling polypeptide, a functional fragment orvariant thereof or a pharmaceutically acceptable salt thereof.Self-assembling polypeptide are polypeptides capable of undergoingspontaneous assembling into ordered nanostructures. Effectivelyself-assembling polypeptides can act as building blocks to form thescaffold domain of the present disclosure. In some embodiments, theself-assembling polypeptides encoded by the expressible nucleic acidsequence of the disclosure are monomeric forms of viral trimers orvariants thereof. In some embodiments, the self-assembling polypeptidesare monomers of nanoparticle structural proteins that self-assemble intonanoparticles upon expression. Any self-assembling polypeptide can beused. In some embodiments, the self assembling polypeptide is fromAquifex aeolicus, Helicobacter pylori, Pyrococcus furiosus or Thermotogamaritima.

A non-limiting example of a self-assembling polypeptide is the lumazinesynthase of hyperthermophilic bacterium Aquifex aeolicus having theamino acid sequence of SEQ ID NO: 8 (LS scaffold) encoded by the nucleicacid sequence of SEQ ID NO: 7.

(SEQ ID NO: 7) atgcagatctacgaaggaaaactgaccgctgagggactgaggttcggaattgtcgcaagccgcgcgaatcacgcactggtggataggctggtggaaggcgctatcgacgcaattgtccggcacggcgggagagaggaagacatcacactggtgagagtctgcggcagctgggagattcccgtggcagctggagaactggctcgaaaggaggacatcgatgccgtgatcgctattggggtcctgtgccgaggagcaactcccagcttcgactacatcgcctcagaagtgagcaaggggctggctgatctgtccctggagctgaggaaacctatcacttttggcgtgattactgccgacaccctggaacaggcaatcgaggcggccggcacctgccatggaaacaaaggctgggaagcagccctgtgcgctattgagatggcaaatctgttc aaatctctgcga(SEQ ID NO: 8) MQIYEGKLTAEGLRFGIVASRANHALVDRLVEGAIDAIVRHGGREEDITLVRVCGSWEIPVAAGELARKEDIDAVIAIGVLCRGATPSFDYIASEVSKGLADLSLELRKPITFGVITADTLEQAIEAAGTCHGNKGWEAALCAIEMANLF KSLR

Another non-limiting example of a self-assembling polypeptide isferritin from Helicobacter pylori having the amino acid sequence of SEQID NO: 10 (3BVE scaffold) encoded by the nucleic acid sequence of SEQ IDNO: 9.

(SEQ ID NO: 9) gggctgagtaaggacattatcaagctgctgaacgaacaggtgaacaaagagatgcagtctagcaacctgtacatgtccatgagctcctggtgctatacccactctctggacggagcaggcctgttcctgtttgatcacgccgccgaggagtacgagcacgccaagaagctgatcatcttcctgaatgagaacaatgtgcccgtgcagctgacctctatcagcgcccctgagcacaagttcgagggcctgacacagatctttcagaaggcctacgagcacgagcagcacatctccgagtctatcaacaatatcgtggaccacgccatcaagtccaaggatcacgccacattcaactttctgcagtggtacgtggccgagcagcacgaggaggaggtgctgtttaaggacatcctggataagatcgagctgatcggcaatgagaaccacgggctgtacctggcagatcagtatgtcaagggcatcgctaagtcaaggaaaag c (SEQ ID NO: 10)GLSKDIIKLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEEYEHAKKLIIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYEHEQHISESINNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIELIGNENHG LYLADQYVKGIAKSRKS

A yet another non-limiting example of a self-assembling polypeptide isPfV viral cage from Pyrococcus furiosus (2e0z) having the amino acidsequence of SEQ ID NO: 12 (RBE scaffold) encoded by the nucleic acidsequence of SEQ ID NO: 11.

(SEQ ID NO: 11) ctgagcattgcccccacactgattaaccgggacaaaccctacaccaaagaggaactgatggagattctgagactggctattatcgctgagctggacgccatcaacctgtacgagcagatggcccggtattctgaggacgagaatgtgcgcaagatcctgctggatgtggccagggaggagaaggcacacgtgggagagttcatggccctgctgctgaacctggaccccgagcaggtgaccgagctgaagggcggctttgaggaggtgaaggagctgacaggcatcgaggcccacatcaacgacaataagaaggaggagagcaacgtggagtatttcgagaagctgagatccgccctgctggatggcgtgaataagggcaggagcctgctgaagcacctgcctgtgaccaggatcgagggccagagcttcagagtggacatcatcaagtttgaggatggcgtgcgcgtggtgaagcaggagtacaagcccatccctctgctgaagaagaagttctacgtgggcatcagggagctgaacgacggcacctacgatgtgagcatcgccacaaaggccggcgagctgctggtgaaggacgaggagtccctggtcatccgcgagatcctgtctacagagggcatcaagaagatgaagctgagctcctgggacaatccagaggaggccctgaacgatctgatgaatgccctgcaggaggcatctaacgcaagcgccggaccattcggcctgatcatcaatcccaagagatacgccaagctgctgaagatctatgagaagtccggcaagatgctggtggaggtgctgaaggagatcttccggggcggcatcatcgtgaccctgaacatcgatgagaacaaagtgatcatctttgccaacacccctgccgtgctggacgtggtggtgggacaggatgtgacactgcaggagctgggaccagagggcgacgatgtggcctttctggtgtccgaggccatcggcatcaggatcaagaatccagaggcaatcgtggtgctggag (SEQ ID NO: 12)LSIAPTLINRDKPYTKEELMEILRLAIIAELDAINLYEQMARYSEDENVRKILLDVAREEKAHVGEFMALLLNLDPEQVTELKGGFEEVKELTGIEAHINDNKKEESNVEYFEKLRSALLDGVNKGRSLLKHLPVTRIEGQSFRVDIIKFEDGVRVVKQEYKPIPLLKKKFYVGIRELNDGTYDVSIATKAGELLVKDEESLVIREILSTEGIKKMKLSSWDNPEEALNDLMNALQEASNASAGPFGLIINPKRYAKLLKIYEKSGKMLVEVLKEIFRGGIIVTLNIDENKVIIFANTPAVLDVVVGQDVTLQELGPEGDDVAFLVSEAIGIRIKNPEAIVVLE

A further non-limiting example of a self-assembling polypeptide is theself-assembling polypeptide from Thermotoga maritima having the aminoacid sequence of SEQ ID NO: 14 (13 scaffold) encoded by the nucleic acidsequence of SEQ ID NO: 13.

(SEQ ID NO: 13) atgaagatggaagaactgttcaagaagcacaagatcgtggccgtgctgagggccaactccgtggaggaggccaagaagaaggccctggccgtgttcctgggcggcgtgcacctgatcgagatcacctttacagtgcccgacgccgataccgtgatcaaggagctgtctttcctgaaggagatgggagcaatcatcggagcaggaaccgtgacaagcgtggagcagtgcagaaaggccgtggagagcggcgccgagtttatcgtgtcccctcacctggacgaggagatctctcagttctgtaaggagaagggcgtgttttacatgccaggcgtgatgacccccacagagctggtgaaggccatgaagctgggccacacaatcctgaagctgttccctggcgaggtggtgggcccacagtttgtgaaggccatgaagggccccttccctaatgtgaagtttgtgcccaccggcggcgtgaacctggataacgtgtgcgagtggttcaaggcaggcgtgctggcagtgggcgtgggcagcgccctggtgaagggcacacccgtggaagtcgctgagaaggcaaaggcattcgtggaaaagatt agggggtgtactgag(SEQ ID NO: 14) MKMEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVHLIEITFTVPDADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAVESGAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKAMKLGHTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCEWFKAGVLAVGVGSALVKGTPVEVAEKAKAFVEKI RGCTE

Additional non-limiting examples of self-assembling polypeptidesinclude: QB scaffold (SEQ ID NO: 16) encoded by the nucleic acidsequence of SEQ ID NO: 15;

(SEQ ID NO: 15) gcaaagctggagacagtgacactgggcaacatcggcaaggacggcaagcagacactggtgctgaatcccaggggcgtgaaccctaccaatggagtggcatctctgagccaggcaggagcagtgcctgccctggagaagagagtgaccgtgtccgtgtctcagcccagcaggaacagaaagaattataaggtgcaggtgaagatccagaacccaaccgcctgcacagccaatggcagctgtgacccatccgtgacaaggcaggcatacgcagatgtgaccttctcttttacacagtatagcaccgatgaggagagggccttcgtgcgcaccgagctggccgccctgctggcatcccctctgctgattgacgctattgaccagctgaaccctgcttac (SEQ ID NO: 16)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGAVPALEKRVTVSVSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTDEERAFVRTELAALLASPLLIDAIDQLNPAYICI scaffold (SEQ ID NO: 18) encoded by thenucleic acid sequence of SEQ ID NO: 17; (SEQ ID NO: 17)gaccctgagtttaccaaaaatgctctgaatgtcgtcaaaaatgatctgattgctaaggtggaccagctgagcggagagcaggaggtgctgaggggcgagctggaggccgccaagcaggcaaaggtgaaactggaaaaccgaatcaaggaactggaagaagaactgaaaagagtc (SEQ ID NO: 18)DPEFTKNALNVVKNDLIAKVDQLSGEQEVLRGELEAAKQAKVKLENRIKE LEEELKRVand IC2 scaffold (SEQ ID NO: 20) encoded by thenucleic acid sequence of SEQ ID NO: 19. (SEQ ID NO: 19)gccgaccccaagaaggtgctggataaagccaaagatcaggcagaaaatagagtcagggaactgaagcagaagctggaggagctgtacaaggaggcccggaagctggacctgacccaggagatgaggagaaagctggagctgcgctacatcgccgccatgctgatggccatcggcgacatctataacgccatcaggcaggccaagcaggaggccgataagctgaagaaggccggcctggtgaatagccagcagctggacgagctgaagcggcgcctggaggagctgaaggaggaggccagcaggaaggccagagattacggcagggagttccagctgaagctggagtatggcggcggcagcggctccggctctggcggcaagatcgagcagatcctgcagaagatcgaaaagatcctgcagaagattgagtggattctgcagaagattgaa cagatcctgcagggg(SEQ ID NO: 20) ADPKKVLDKAKDQAENRVRELKQKLEELYKEARKLDLTQEMRRKLELRYIAAMLMAIGDIYNAIRQAKQEADKLKKAGLVNSQQLDELKRRLEELKEEASRKARDYGREFQLKLEYGGGSGSGSGGKIEQILQKIEKILQKIEWILQKIE QILQG

Accordingly, in some embodiments, the self-assembling polypeptideencoded by the expressible nucleic acid sequence of the presentdisclosure comprises at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%sequence identity to SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ IDNO: 14, SEQ ID NO: 16, SEQ ID NO: 18 or SEQ ID NO: 20, or a functionalfragment or variant thereof. In some embodiments, the self-assemblingpolypeptide comprises the amino acid sequence of SEQ ID NO: 8, SEQ IDNO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18 orSEQ ID NO: 20, or a functional fragment or variant thereof. In someembodiments, the nucleic acid sequence encoding the self-assemblingpolypeptide comprises at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%sequence identity to SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ IDNO: 13, SEQ ID NO: 15, SEQ ID NO: 17 or SEQ ID NO: 19, or a functionalfragment or variant thereof. In some embodiments, the nucleic acidsequence encoding the self-assembling polypeptide comprises thenucleotide sequence of SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ IDNO: 13, SEQ ID NO: 15, SEQ ID NO: 17 or SEQ ID NO: 19, or a functionalfragment or variant thereof. In other embodiments, the nucleic acidsequence encoding the self-assembling polypeptide comprises at leastabout 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a nucleic acidsequence that is complementary to SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO:11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17 or SEQ ID NO: 19, or afunctional fragment thereof. In some embodiments, the nucleic acidsequence encoding the self-assembling polypeptide comprises a nucleicacid sequence that is complementary to SEQ ID NO: 7, SEQ ID NO: 9, SEQID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17 or SEQ ID NO: 19,or a functional fragment thereof.

In some embodiments, the composition or pharmaceutical composition ofthe disclosure comprises a vector or a nucleic acid molecule comprisingan expressible RNA sequence that encodes a self-assembling polypeptidethat is optionally in sequence with one or more additional expressibleRNA sequences that encode a viral antigen. In some embodiments, theexpressible DNA or RNA sequence that encodes a self-assemblingpolypeptide is from about 300 to about 500 nucleotides in length. Insome embodiments, the expressible DNA or RNA sequence that encodes aself-assembling polypeptide is from about 350 to about 480 nucleotidesin length. In some embodiments, the expressible DNA or RNA sequence thatencodes a self-assembling polypeptide is from about 350 to about 460nucleotides in length. In some embodiments, the expressible DNA or RNAsequence that encodes a self-assembling polypeptide is from about 300 toabout 500 nucleotides in length. In some embodiments, the expressibleDNA or RNA sequence that encodes a self-assembling polypeptide is fromabout 400 to about 500 nucleotides in length. In some embodiments, theexpressible DNA or RNA sequence that encodes a self-assemblingpolypeptide is from about 390 to about 410 nucleotides in length. Insome embodiments, the expressible DNA or RNA sequence that encodes aself-assembling polypeptide is from about 300 to about 410 nucleotidesin length. In some embodiments, the expressible DNA or RNA sequence thatencodes a self-assembling polypeptide is from about 300 to about 500nucleotides in length.

3. Linker

The expressible nucleic acid sequence of the present disclosureoptionally comprises a nucleic acid sequence encoding a linker peptide,a functional fragment or variant thereof or a pharmaceuticallyacceptable salt thereof. Any type of linker or linker peptide can beused. The term “linker” or “linker peptide” is used interchangeableherein.

In some embodiments, each linker or linker peptide is independentlyselectable from about 0 to about 25, about 1 to about 25, about 2 toabout 25, about 3 to about 25, about 4 to about 25, about 5 to about 25,about 6 to about 25, about 7 to about 25, about 8 to about 25, about 9to about 25, about 10 to about 25, about 11 to about 25, about 12 toabout 25, about 13 to about 25, about 14 to about 25, about 15 to about25, about 16 to about 25, about 17 to about 25, about 18 to about 25,about 19 to about 25, about 20 to about 25, about 21 to about 25, about22 to about 25, about 23 to about 25, about 24 to about 25 natural ornon-natural amino acids in length.

In some embodiments, each linker or linker peptide is about 0, about 1,about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9,about 10, about 11, about 12, about 13, about 14, about 15, about 16,about 17, about 18, about 19, about 20, about 21, about 22, about 23,about 24, about 25 natural or non-natural amino acids in length. In someembodiments, each linker or linker peptide is independently selectablefrom a linker or linker peptide that is about 0, about 1, about 2, about3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about11, about 12, about 13, about 14, about 15, about 16, about 17, about18, about 19, about 20, about 21, about 22, about 23, about 24, about 25natural or non-natural amino acids in length. In some embodiments, eachlinker or linker peptide is about 21 natural or non-natural amino acidsin length.

In some embodiments, the length of each linker or linker peptide isdifferent. For example, in some embodiments, the length of a firstlinker or linker peptide is about 0, about 1, about 2, about 3, about 4,about 5, about 6, about 7, about 8, about 9, about 10, about 11, about12, about 13, about 14, about 15, about 16, about 17, about 18, about19, about 20, about 21, about 22, about 23, about 24, about 25 naturalor non-natural amino acids in length, and the length of a second linkeris about 0, about 1, about 2, about 3, about 4, about 5, about 6, about7, about 8, about 9, about 10, about 11, about 12, about 13, about 14,about 15, about 16, about 17, about 18, about 19, about 20, about 21,about 22, about 23, about 24, about 25 natural or non-natural aminoacids in length, where the length of the first linker is different fromthe length of the second linker. Various configurations can beenvisioned by the present disclosure, where the linker domain comprises1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more linkers or linker peptides whereinthe linkers or linker peptides are of similar or different lengths.

In some embodiments, two linkers or linker peptides can be usedtogether. Accordingly, in some embodiments, the first linker or linkerpeptide is independently selectable from about 0 to about 25 natural ornon-natural amino acids in length, about 0 to about 25, about 1 to about25, about 2 to about 25, about 3 to about 25, about 4 to about 25, about5 to about 25, about 6 to about 25, about 7 to about 25, about 8 toabout 25, about 9 to about 25, about 10 to about 25, about 11 to about25, about 12 to about 25, about 13 to about 25, about 14 to about 25,about 15 to about 25, about 16 to about 25, about 17 to about 25, about18 to about 25, about 19 to about 25, about 20 to about 25, about 21 toabout 25, about 22 to about 25, about 23 to about 25, about 24 to about25 natural or non-natural amino acids in length. In some embodiments,the second linker or linker peptide is independently selectable fromabout 0 to about 25, about 1 to about 25, about 2 to about 25, about 3to about 25, about 4 to about 25, about 5 to about 25, about 6 to about25, about 7 to about 25, about 8 to about 25, about 9 to about 25, about10 to about 25, about 11 to about 25, about 12 to about 25, about 13 toabout 25, about 14 to about 25, about 15 to about 25, about 16 to about25, about 17 to about 25, about 18 to about 25, about 19 to about 25,about 20 to about 25, about 21 to about 25, about 22 to about 25, about23 to about 25, about 24 to about 25 natural or non-natural amino acidsin length. In some embodiments, the first linker or linker peptide isindependently selectable from a linker or linker peptide that is about0, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about8, about 9, about 10, about 11, about 12, about 13, about 14, about 15,about 16, about 17, about 18, about 19, about 20, about 21, about 22,about 23, about 24, about 25 natural or non-natural amino acids inlength. In some embodiments, the second linker or linker peptide isindependently selectable from a linker or linker peptide that is about0, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about8, about 9, about 10, about 11, about 12, about 13, about 14, about 15,about 16, about 17, about 18, about 19, about 20, about 21, about 22,about 23, about 24, about 25 natural or non-natural amino acids inlength.

A non-limiting example of a linker peptide may comprise the amino acidsequence of GGSGGSGGSGGSGGG (SEQ ID NO: 22) encoded by the nucleic acidsequence of ggaggctccggaggatctggagggagtggaggctcaggaggaggc (SEQ ID NO:21).

A linker or linker peptide can be either flexible or rigid or acombination thereof. An example of a flexible linker is a GGS repeat. Insome embodiments, the GGS can be repeated about 1, 2, 3, 4, 5, 6, 7, 8,9, or 10 times. Non-limiting examples of such linker peptides maycomprise the amino acid sequence of GGSGGSGGS (SEQ ID NO: 23),GGSGGSGGSGGS (SEQ ID NO: 24), or GGSGGSGGSGGSGGGGSGGGSGGG (SEQ ID NO:25). An example of a rigid linker is 4QTL-115 Angstroms, single chain3-helix bundle represented by the sequence:

(SEQ ID NO: 26) NEDDMKKLYKQMVQELEKARDRMEKLYKEMVELIQKAIELMRKIFQEVKQEVEKAIEEMKKLYDEAKKKIEQMIQQIKQGGDKQKMEELLKRAKEEMKKVKDKMEKLLEKLKQIMQEAKQKMEKLLKQLKEEMKKMKEKMEKLLKEMKQRMEEVKKKMDGDDELLEKIKKNIDDLKKIAEDLIKKAEENIKEAKKIAEQLVKRAKQLIEKAKQVAEELIKKILQLIEKAKEIAEKVLKGLE.Other non-limiting examples of linker peptides may be encoded by the nucleicacid sequence of (SEQ ID NO: 27)ggcggctctggcggaagtggcggaagtgggggaagtggaggcggcggaagcgggggaggcagcgggggaggg,(SEQ ID NO: 28) ggcggaagcggcggaagcggcgggtct, (SEQ ID NO: 29)ggcggcagcggcggcagcggcgggagcggaggaagt, or (SEQ ID NO: 30)ggcggctctggcggaagtggcggaagtgggggaagtggaggcggcggaagcgggggaggcagcgggggaggg.Additional non-limiting examples of linker peptides include Link 14 linker (SEQID NO: 32) encoded by the nucleic acid sequence of SEQ ID NO: 31;(SEQ ID NO: 31) tctcacagcggctccggcggctctggcagcggcggccacgcc(SEQ ID NO: 32) SHSGSGGSGSGGHACPG9.2 linker 1 (SEQ ID NO: 34) encoded by the nucleic acid sequence of SEQ ID NO: 33;(SEQ ID NO: 33) gggggaaatagtagcggc (SEQ ID NO: 34) GGNSSGCPG9.2 linker 2 (SEQ ID NO: 36) encoded by the nucleic acid sequence of SEQ ID NO: 35;(SEQ ID NO: 35) ggcggcaacggcagcggcggcggcagcggctccggcggcaacggctctagcggc(SEQ ID NO: 36) GGNGSGGGSGSGGNGSSGPDGFR linker (between tri mer or TSI and PDGFR; SEQ ID NO: 38) encoded by the nucleicacid sequence of SEQ ID NO: 37; (SEQ ID NO: 37)ggaggaggaagcgggggaagcgggggaagcggaggaagcgggggaagcgggggaagc(SEQ ID NO: 38) GGGSGGSGGSGGSGGSGGSFoldon PDGFR linker 1 (SEQ ID NO: 40) encoded by the nucleic acid sequence of SEQ IDNO: 39; (SEQ ID NO: 39) ggaggaggaagcgggggaagcggcggcggc (SEQ ID NO: 40)GGGSGGSGGGFoldon PDGFR linker 2 (SEQ ID NO: 42) encoded by the nucleic acid sequence of SEQ IDNO: 41; (SEQ ID NO: 41) gggggaagcggaggaagcgggggaagcgggggaagc(SEQ ID NO: 42) GGSGGSGGSGGS3BVE linker (SEQ ID NO: 44) encoded by the nucleic acid sequence of SEQ ID NO: 43;(SEQ ID NO: 43) ggaagcggc (SEQ ID NO: 44) GSG13_1 linker (SEQ ID NO: 46) encoded by the nucleic acid sequence of SEQ ID NO: 45;(SEQ ID NO: 45) ggcggcagcggcagcggcgggagcggagga (SEQ ID NO: 46)GGSGSGGSGG13_2 linker (SEQ ID NO: 48) encoded by the nucleic acid sequence of SEQ ID NO: 47;(SEQ ID NO: 47)ggagggagcgatatgagaaaggacgccgagagacggtttgataagttcgtggaggctgctaagaataagtttgacaagtttaaggctgccctgcggaagggcgacatcaaggaggagaggagaaaggatatgaagaagctggcaaggaaggaggcagagcaggcaaggagggccgtgaggaacagactgagcgagctgctgtccaagatcaacgacatgcccatcaccaatgatcagaagaagctgatgtctaatgacgtgctgaagttcgccgcagaagccgaaaagaagattgaagccctggcagcagacgccgaaggaggaagcgggagc(SEQ ID NO: 48) GGSDMRKDAERRFDKFVEAAKNKFDKFKAALRKGDIKEERRKDMKKLARKEAEQARRAVRNRLSELLSKINDMPITNDQKKLMSNDVLKFAAEAEKKIEALAADAEGGSGSLS_1 linker (SEQ ID NO: 50) encoded by the nucleic acid sequence of SEQ ID NO: 49;(SEQ ID NO: 49)gggggctctagcgggaaaagtctggtggataccgtctatgctctgaaagatgaggtgcaggaactgaggcaggacaacaaaaagatgaagaagagcctggaggaggagcagagggccagaaaggacctggaaaaactggtgcggaaagtgctgaaaaacatgaatgacggagggagtagcggg (SEQ ID NO: 50)GGSSGKSLVDTVYALKDEVQELRQDNKKMKKSLEEEQRARKDLEKLVRKVLKNMN DGGSSGLS_2 linker (SEQ ID NO: 52) encoded by the nucleic acid sequence of SEQ ID NO: 51;(SEQ ID NO: 51)gggggctctagcggggcagacccaaagaaagtgctggataaggcaaaggatcaggcagagaatagagtgagagaactgaaacagaaactggaggaactgtataaggaggcccggaagctggacctgacccaggagatgaggagaaagctggagctgcgctacatcgccgccatgctgatggccatcggcgacatctataacgccatcaggcaggccaagcaggaggccgataagctgaagaaggccggcctggtgaatagccagcagctggacgagctgaagcggcgcctggaggagctgaaggaggaggcctccaggaaggccagagattatgggcgggaatttcagctgaaactggagtatggcggcggaagcggaagcgggagcggg (SEQ ID NO: 52)GGSSGADPKKVLDKAKDQAENRVRELKQKLEELYKEARKLDLTQEMRRKLELRYIAAMLMAIGDIYNAIRQAKQEADKLKKAGLVNSQQLDELKRRLEELKEEASRKARDYGREFQLKLEYGGGSGSGSGQB_1 linker (SEQ ID NO: 54) encoded by the nucleic acid sequence of SEQ ID NO: 53;(SEQ ID NO: 53)ggaggctcttcaggcggcacagacgtgggggcaatcgctggaaaggctaacgaggctggacagggggcttatgatgctcaggtcaaaaacgacgagcaggatgtggagctggccgaccacgaggccaggatcaagcagctgagaatcgatgtggacgatcacgagtctcggatcaccgccaacacaaaggccatcacagccctgaatgtgcgcgtgaccacagcagagggagagatcgcatccctgcagaccaacgtgagcgccctggacggaagggtgaccacagcagagaacaatatctccgccctgcaggcagattacgtgagcggcggcagctccggctccgga (SEQ ID NO: 54)GGSSGGTDVGAIAGKANEAGQGAYDAQVKNDEQDVELADHEARIKQLRIDVDDHESRITANTKAITALNVRVTTAEGEIASLQTNVSALDGRVTTAENNISALQADYVSGGSS GSGQB 2 linker (SEQ ID NO: 56) encoded by the nucleic acid sequence of SEQ ID NO: 55;and (SEQ ID NO: 55)ggaggctctggaagcgggggaagtagcggacctcacatgattgctccaggacatcgggacgagtttgaccctaagctgccaacaggcgagaaagaagaggtgccaggcaagcccggcatcaagaaccctgagacaggcgacgtggtgaggccccctgtggattctgtgacaaagtacggcccagtgaagggcgacagcatcgtggagaaggaggagatccccttcgagaaggagaggaagtttaaccctgatctggccccaggcaccgagaaggtgacaagagagggccagaagggcgagaagaccatcaccacacccacactgaagaatcctctgaccggcgagatcatcagcaagggcgagtccaaggaggagatcacaaaggaccccatcaacgaactgaccgaatggggaccagagacaggaggaagcggcagcggcggaagcagc (SEQ ID NO: 56)GGSGSGGSSGPHMIAPGHRDEFDPKLPTGEKEEVPGKPGIKNPETGDVVRPPVDSVTKYGPVKGDSIVEKEEIPFEKERKFNPDLAPGTEKVTREGQKGEKTITTPTLKNPLTGEIISKGESKEEITKDPINELTEWGPETGGSGSGGSSIC1/IC2 linker (SEQ ID NO: 58) encoded by the nucleic acid sequence of SEQ ID NO: 57(SEQ ID NO: 57) ggaggcagcggcagcggcagcggg (SEQ ID NO: 58) GGSGSGSG

Accordingly, in some embodiments, the linker peptide encoded by theexpressible nucleic acid sequence of the present disclosure comprises atleast about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ IDNO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50,SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56 or SEQ ID NO: 58, or afunctional fragment or variant thereof. In some embodiments, the linkerpeptide comprises the amino acid sequence of SEQ ID NO: 22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 32, SEQ IDNO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52,SEQ ID NO: 54, SEQ ID NO: 56 or SEQ ID NO: 58, or a functional fragmentor variant thereof. In some embodiments, the nucleic acid sequenceencoding the linker peptide comprises at least about 60%, 65%, 70%, 75%,80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% sequence identity to SEQ ID NO: 21, SEQ ID NO: 27, SEQ ID NO:28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 33, SEQ IDNO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53,SEQ ID NO: 55 or SEQ ID NO: 57 or a functional fragment or variantthereof. In some embodiments, the nucleic acid sequence encoding thelinker peptide comprises the nucleotide sequence of SEQ ID NO: 21, SEQID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31,SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO:41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ IDNO: 51, SEQ ID NO: 53, SEQ ID NO: 55 or SEQ ID NO: 57 or a functionalfragment or variant thereof. In other embodiments, the nucleic acidsequence encoding the linker peptide comprises at least about 60%, 65%,70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98% or 99% sequence identity to a nucleic acid sequence thatis complementary to SEQ ID NO: 21, SEQ ID NO: 27, SEQ ID NO: 28, SEQ IDNO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45,SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO:55 or SEQ ID NO: 57 or a functional fragment or variant thereof. In someembodiments, the nucleic acid sequence encoding the linker peptidecomprises a nucleic acid sequence that is complementary to thenucleotide sequence of SEQ ID NO: 21, SEQ ID NO: 27, SEQ ID NO: 28, SEQID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35,SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO:45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ IDNO: 55 or SEQ ID NO: 57 or a functional fragment or variant thereof

4. Viral Antigens

The expressible nucleic acid sequence of the present disclosurecomprises a nucleic acid sequence encoding an antigen domain comprisinga viral antigen, a functional fragment or variant thereof or apharmaceutically acceptable salt thereof. In some embodiments, the viralantigen comprises an antigen from a virus from the family ofCoronaviridae. In some embodiments, the viral antigen comprises anantigen from a coronavirus. In some embodiments, the viral antigencomprises an antigen from SARS-CoV. In some embodiments, the viralantigen comprises an antigen from SARS-CoV-2. In some embodiments, theviral antigen comprises the spike protein of SARS-CoV-2, a functionalfragment or variant thereof or a pharmaceutically acceptable saltthereof. In some embodiments, the viral antigen comprises a viral trimerpolypeptide, a functional fragment or variant thereof or apharmaceutically acceptable salt thereof. In some embodiments, the viraltrimer comprises a trimer from a virus from the family of Coronaviridae.In some embodiments, the viral trimer comprises a trimer from acoronavirus. In some embodiments, the viral trimer comprises a trimerfrom SARS-CoV. In some embodiments, the viral trimer comprises a trimerfrom SARS-CoV-2. In some embodiments, the viral trimer comprises thespike protein of SARS-CoV-2, a functional fragment or variant thereof ora pharmaceutically acceptable salt thereof.

A non-limiting example of a viral antigen is a fragment of the surfaceglycoprotein (or spike protein or S protein) of SARS-CoV-2 having theamino acid sequence of SEQ ID NO: 60 encoded by the nucleic acidsequence of SEQ ID NO: 59 (GenBank Accession No. QHD43416).

(SEQ ID NO: 59)atgtttgtttttcttgttttattgccactagtctctagtcagtgtgttaatcttacaaccagaactcaattaccccctgcatacactaattctttcacacgtggtgtttattaccctgacaaagttttcagatcctcagttttacattcaactcaggacttgttcttacctttcttttccaatgttacttggttccatgctatacatgtctctgggaccaatggtactaagaggtttgataaccctgtcctaccatttaatgatggtgtttattttgcttccactgagaagtctaacataataagaggctggatttttggtactactttagattcgaagacccagtccctacttattgttaataacgctactaatgttgttattaaagtctgtgaatttcaattttgtaatgatccatttttgggtgtttattaccacaaaaacaacaaaagttggatggaaagtgagttcagagtttattctagtgcgaataattgcacttttgaatatgtctctcagccttttcttatggaccttgaaggaaaacagggtaatttcaaaaatcttagggaatttgtgtttaagaatattgatggttattttaaaatatattctaagcacacgcctattaatttagtgcgtgatctccctcagggtttttcggctttagaaccattggtagatttgccaataggtattaacatcactaggtttcaaactttacttgctttacatagaagttatttgactcctggtgattcttcttcaggttggacagctggtgctgcagcttattatgtgggttatcttcaacctaggacttttctattaaaatataatgaaaatggaaccattacagatgctgtagactgtgcacttgaccctctctcagaaacaaagtgtacgttgaaatccttcactgtagaaaaaggaatctatcaaacttctaactttagagtccaaccaacagaatctattgttagatttcctaatattacaaacttgtgcccttttggtgaagtttttaacgccaccagatttgcatctgtttatgcttggaacaggaagagaatcagcaactgtgttgctgattattctgtcctatataattccgcatcattttccacttttaagtgttatggagtgtctcctactaaattaaatgatctctgctttactaatgtctatgcagattcatttgtaattagaggtgatgaagtcagacaaatcgctccagggcaaactggaaagattgctgattataattataaattaccagatgattttacaggctgcgttatagcttggaattctaacaatcttgattctaaggttggtggtaattataattacctgtatagattgtttaggaagtctaatctcaaaccttttgagagagatatttcaactgaaatctatcaggccggtagcacaccttgtaatggtgttgaaggttttaattgttactttcctttacaatcatatggtttccaacccactaatggtgttggttaccaaccatacagagtagtagtactttcttttgaacttctacatgcaccagcaactgtttgtggacctaaaaagtctactaatttggttaaaaacaaatgtgtcaatttcaacttcaatggtttaacaggcacaggtgttcttactgagtctaacaaaaagtttctgcctttccaacaatttggcagagacattgctgacactactgatgctgtccgtgatccacagacacttgagattcttgacattacaccatgttcttttggtggtgtcagtgttataacaccaggaacaaatacttctaaccaggttgctgttctttatcaggatgttaactgcacagaagtccctgttgctattcatgcagatcaacttactcctacttggcgtgtttattctacaggttctaatgtttttcaaacacgtgcaggctgtttaataggggctgaacatgtcaacaactcatatgagtgtgacatacccattggtgcaggtatatgcgctagttatcagactcagactaattctcctcggcgggcacgtagtgtagctagtcaatccatcattgcctacactatgtcacttggtgcagaaaattcagttgcttactctaataactctattgccatacccacaaattttactattagtgttaccacagaaattctaccagtgtctatgaccaagacatcagtagattgtacaatgtacatttgtggtgattcaactgaatgcagcaatcttttgttgcaatatggcagtttttgtacacaattaaaccgtgctttaactggaatagctgttgaacaagacaaaaacacccaagaagtttttgcacaagtcaaacaaatttacaaaacaccaccaattaaagattttggtggttttaatttttcacaaatattaccagatccatcaaaaccaagcaagaggtcatttattgaagatctacttttcaacaaagtgacacttgcagatgctggcttcatcaaacaatatggtgattgccttggtgatattgctgctagagacctcatttgtgcacaaaagtttaacggccttactgttttgccacctttgctcacagatgaaatgattgctcaatacacttctgcactgttagcgggtacaatcacttctggttggacctttggtgcaggtgctgcattacaaataccatttgctatgcaaatggcttataggtttaatggtattggagttacacagaatgttctctatgagaaccaaaaattgattgccaaccaatttaatagtgctattggcaaaattcaagactcactttcttccacagcaagtgcacttggaaaacttcaagatgtggtcaaccaaaatgcacaagctttaaacacgcttgttaaacaacttagctccaattttggtgcaatttcaagtgttttaaatgatatcctttcacgtcttgacaaagttgaggctgaagtgcaaattgataggttgatcacaggcagacttcaaagtttgcagacatatgtgactcaacaattaattagagctgcagaaatcagagcttctgctaatcttgctgctactaaaatgtcagagtgtgtacttggacaatcaaaaagagttgatttttgtggaaagggctatcatcttatgtccttccctcagtcagcacctcatggtgtagtcttcttgcatgtgacttatgtccctgcacaagaaaagaacttcacaactgctcctgccatttgtcatgatggaaaagcacactttcctcgtgaaggtgtctttgtttcaaatggcacacactggtttgtaacacaaaggaatttttatgaaccacaaatcattactacagacaacacatttgtgtctggtaactgtgatgttgtaataggaattgtcaacaacacagtttatgatcctttgcaacctgaattagactcattcaaggaggagttagataaatattttaagaatcatacatcaccagatgttgatttaggtgacatctctggcattaatgcttcagttgtaaacattcaaaaagaaattgaccgcctcaatgaggttgccaagaatttaaatgaatctctcatcgatctccaagaacttggaaagtatgagcagtatataaaatggccatggtacatttggctaggttttatagctggcttgattgccatagtaatggtgacaattatgctttgctgtatgaccagttgctgtagttgtctcaagggctgttgttcttgtggatcctgctgcaaatttgatgaagacgactctgagccagtgctcaaaggagtcaaattacattacacataa(SEQ ID NO: 60)MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGV KLHYT

Non-limiting examples of fragments of the S protein of SARS-CoV-2comprises the following sequences:

(SEQ ID NO: 171)MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIK WPW(SEQ ID NO: 172)MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIK WP(SEQ ID NO: 173)MFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQ (SEQ ID NO: 174)SQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQ (SEQ ID NO: 175)NLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDIST (SEQ ID NO: 176)SFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKF (SEQ ID NO: 177)PSKRSFIEDLLFNKV

A further non-limiting example of a viral antigen is a fragment of theenvelop protein (or E protein) of SARS-CoV-2 having the amino acidsequence of SEQ ID NO: 62 encoded by the nucleic acid sequence of SEQ IDNO: 61 (GenBank Accession No. QHD43418).

(SEQ ID NO: 61) atgtactcattcgtttcggaagagacaggtacgttaatagttaatagcgtacttctttttcttgctttcgtggtattcttgctagttacactagccatccttactgcgcttcgattgtgtgcgtactgctgcaatattgttaacgtgagtcttgtaaaaccttctttttacgtttactctcgtgttaaaaatctgaattcttctagagttcctgatcttctggtctaa (SEQ ID NO: 62)MYSFVSEETGTLIVNSVLLFLAFVVFLLVTLAILTALRLCAYCCNIVNVSLVKPSFYVYSRVKNLNSSRVPDLLV

Another non-limiting example of a viral antigen is a fragment of themembrane glycoprotein (or M protein) of SARS-CoV-2 having the amino acidsequence of SEQ ID NO: 64 encoded by the nucleic acid sequence of SEQ IDNO: 63 (GenBank Accession No. QHD43419).

(SEQ ID NO: 63) ccatggcagattccaacggtactattaccgttgaagagcttaaaaagctccttgaacaatggaacctagtaataggtttcctattccttacatggatttgtcttctacaatttgcctatgccaacaggaataggtttttgtatataattaagttaattttcctctggctgttatggccagtaactttagcttgttttgtgcttgctgctgtttacagaataaattggatcaccggtggaattgctatcgcaatggcttgtcttgtaggcttgatgtggctcagctacttcattgcttctttcagactgtttgcgcgtacgcgttccatgtggtcattcaatccagaaactaacattcttctcaacgtgccactccatggcactattctgaccagaccgcttctagaaagtgaactcgtaatcggagctgtgatccttcgtggacatcttcgtattgctggacaccatctaggacgctgtgacatcaaggacctgcctaaagaaatcactgttgctacatcacgaacgctttcttattacaaattgggagcttcgcagcgtgtagcaggtgactcaggttttgctgcatacagtcgctacaggattggcaactataaattaaacacagaccattccagtagcagtgacaat attgctttgcttgtacagtaa(SEQ ID NO: 64) MADSNGTITVEELKKLLEQWNLVIGFLFLTWICLLQFAYANRNRFLYIIKLIFLWLLWPVTLACFVLAAVYRINWITGGIAIAMACLVGLMWLSYFIASFRLFARTRSMWSFNPETNILLNVPLHGTILTRPLLESELVIGAVILRGHLRIAGHHLGRCDIKDLPKEITVATSRTLSYYKLGASQRVAGDSGFAAYSRYRIGNYKLNTDHSSSSDNIALLVQ

A yet another non-limiting example of a viral antigen is a fragment ofthe nucleocapsid phosphoprotein (or N protein) of SARS-CoV-2 having theamino acid sequence of SEQ ID NO: 66 encoded by the nucleic acidsequence of SEQ ID NO: 65 (GenBank Accession No. QHD43423).

(SEQ ID NO: 65) atgtctgataatggaccccaaaatcagcgaaatgcaccccgcattacgtttggtggaccctcagattcaactggcagtaaccagaatggagaacgcagtggggcgcgatcaaaacaacgtcggccccaaggtttacccaataatactgcgtcttggttcaccgctctcactcaacatggcaaggaagaccttaaattccctcgaggacaaggcgttccaattaacaccaatagcagtccagatgaccaaattggctactaccgaagagctaccagacgaattcgtggtggtgacggtaaaatgaaagatctcagtccaagatggtatttctactacctaggaactgggccagaagctggacttccctatggtgctaacaaagacggcatcatatgggttgcaactgagggagccttgaatacaccaaaagatcacattggcacccgcaatcctgctaacaatgctgcaatcgtgctacaacttcctcaaggaacaacattgccaaaaggcttctacgcagaagggagcagaggcggcagtcaagcctcttctcgttcctcatcacgtagtcgcaacagttcaagaaattcaactccaggcagcagtaggggaacttctcctgctagaatggctggcaatggcggtgatgctgctcttgctttgctgctgcttgacagattgaaccagcttgagagcaaaatgtctggtaaaggccaacaacaacaaggccaaactgtcactaagaaatctgctgctgaggcttctaagaagcctcggcaaaaacgtactgccactaaagcatacaatgtaacacaagctttcggcagacgtggtccagaacaaacccaaggaaattttggggaccaggaactaatcagacaaggaactgattacaaacattggccgcaaattgcacaatttgcccccagcgcttcagcgttcttcggaatgtcgcgcattggcatggaagtcacaccttcgggaacgtggttgacctacacaggtgccatcaaattggatgacaaagatccaaatttcaaagatcaagtcattttgctgaataagcatattgacgcatacaaaacattcccaccaacagagcctaaaaaggacaaaaagaagaaggctgatgaaactcaagccttaccgcagagacagaagaaacagcaaactgtgactcttcttcctgctgcagatttggatgatttctccaaacaattgcaacaatccatgagcagtgctgactcaac tcaggcctaa(SEQ ID NO: 66) MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGLPNNTASWFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSRNSTPGSSRGTSPARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADL DDFSKQLQQSMSSADSTQA

Accordingly, in some embodiments, the viral antigen encoded by theexpressible nucleic acid sequence of the present disclosure comprises atleast about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO:60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ IDNO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176or SEQ ID NO: 177, or a functional fragment or variant thereof or apharmaceutically acceptable salt thereof. In some embodiments, the viralantigen comprises the amino acid sequence of SEQ ID NO: 60, SEQ ID NO:62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO: 172, SEQ IDNO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 or SEQ ID NO:177, or a functional fragment or variant thereof or a pharmaceuticallyacceptable salt thereof. In some embodiments, the nucleic acid sequenceencoding the viral antigen comprises at least about 60%, 65%, 70%, 75%,80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% sequence identity to SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO:63 or SEQ ID NO: 65, or a functional fragment or variant thereof or apharmaceutically acceptable salt thereof. In some embodiments, thenucleic acid sequence encoding the viral antigen comprises thenucleotide sequence of SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63 orSEQ ID NO: 65, or a functional fragment or variant thereof or apharmaceutically acceptable salt thereof. In some embodiments, thenucleic acid sequence encoding the viral antigen comprises at leastabout 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a nucleic acidsequence that is complementary to SEQ ID NO: 59, SEQ ID NO: 61, SEQ IDNO: 63 or SEQ ID NO: 65, or a functional fragment or variant thereof ora pharmaceutically acceptable salt thereof. In some embodiments, thenucleic acid sequence encoding the viral antigen comprises a nucleicacid sequence that is complementary to the nucleotide sequence of SEQ IDNO: 59, SEQ ID NO: 61, SEQ ID NO: 63 or SEQ ID NO: 65, or a functionalfragment or variant thereof or a pharmaceutically acceptable saltthereof.

In some embodiments, the composition or pharmaceutical composition ofthe disclosure comprises an expressible DNA or RNA sequence that encodesa viral antigen and is from about 1100 to about 1300 nucletides inlength. In some embodiments, the composition or pharmaceuticalcomposition of the disclosure comprises an expressible DNA or RNAsequence that encodes a viral antigen and is from about 1100 to about1200 nucletides in length. In some embodiments, the composition orpharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1100to about 1300 nucletides in length. In some embodiments, the compositionor pharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1200to about 1210 nucletides in length. In some embodiments, the compositionor pharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1180to about 1220 nucletides in length. In some embodiments, the compositionor pharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1180to about 1215 nucletides in length. In some embodiments, the compositionor pharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1180to about 1210 nucletides in length. In some embodiments, the compositionor pharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1180to about 1200 nucletides in length. In some embodiments, the compositionor pharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1220to about 1230 nucletides in length. In some embodiments, the compositionor pharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1100to about 1300 nucletides in length. In some embodiments, the compositionor pharmaceutical composition of the disclosure comprises an expressibleDNA or RNA sequence that encodes a viral antigen and is from about 1100to about 1300 nucletides in length.

In some embodiments, the expressible nucleic acid sequence encodes afusion protein comprising one or a plurality of coronaviral envelopepolypeptides or functional fragments thereof. In some embodiments, thefusion protein comprise a furin cleavage site. In some embodiments, theexpressible nucleic acid sequence comprises a first nucleic acidsequence encoding, in a 5′ to 3′ orientation, at least three monomers ofcoronaviral envelope proteins. In some embodiments, the at least threemonomers of coronaviral envelope proteins are separated by a furincleavage site. In some embodiments, the furin cleavage site comprises atleast about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to RRRRRR(SEQ ID NO: 67), or a functional fragment or variant thereof or apharmaceutically acceptable salt thereof. In some embodiments, the furincleavage site comprises the amino acid sequence of SEQ ID NO: 67, or afunctional fragment or variant thereof or a pharmaceutically acceptablesalt thereof. In some embodiments, the expressible nucleic acid sequenceencodes a polypeptide free of carbohydrate proximate to at least 30amino acids from the carboxy end of the polypeptide. In someembodiments, the expressible nucleic acid sequence encodes a polypeptidefree of carbohydrate proximate to at least 20 amino acids from thecarboxy end of the polypeptide. In some embodiments, the expressiblenucleic acid sequence encodes a polypeptide free of carbohydrateproximate to at least 10 amino acids from the carboxy end of thepolypeptide. In some embodiments, the expressible nucleic acid sequenceencodes a polypeptide free of carbohydrate proximate to at least 50amino acids from the carboxy end of the polypeptide.

In some embodiments, the expressible nucleic acid sequence of thedisclosure comprises at least a first nucleic acid sequence encoding afirst, a second and/or a third polypeptides, each first, second or thirdpolypeptide comprising a viral antigen. In some embodiments, theexpressible nucleic acid sequence encodes one or a plurality of fusionproteins, each fusion protein comprising at least a first, a second,and/or a third polypeptide contiguously linked by a linker sequence. Insome embodiments, the expressible nucleic acid sequence of thedisclosure comprises at least a first nucleic acid sequence encoding atleast one self-assembling polypeptide. In some embodiments, theself-assembling polypeptide is at least one self-assembling component ofa nanoparticle or at least one coronaviral monomer, the coronaviralmonomer capable of assembling into a coronaviral trimer upon expressionin a cell. In some embodiments, the expressible nucleic acid sequencecomprises a nucleic acid sequence encoding a coronaviral antigen, butfree of a nucleic acid sequence encoding a self-assembling polypeptide.In some embodiments, the expressible nucleic acid sequence of thedisclosure comprises a nucleic acid sequence operably linked to aregulatory sequence and encodes a fusion peptide comprising one or aplurality of self-assembling polypeptides, wherein at least one of theself-assembling polypeptides is a self-assembling coronaviral antigen.

In some embodiments, upon administration to a subject a compositioncomprising the expressible nucleic acid sequence of the disclosure, theexpressible nucleic acid sequence is transfected or transduced into anantigen presenting cell. After a plurality of expressible nucleic acidsequences are expressed, the self-assembling polypeptides assemble withinto a non-native form of a viral antigen. In some embodiments, thenon-native form of a viral antigen comprises a coronaviral trimerexposing an amino acid sequence that is not naturally exposed or free ofcarbohydrate as compared to its corresponding native form or variantsthereof. Expression and presentation of the one or plurality ofself-assembling polypeptides elicits an immune response against anepitope. In some embodiments, the epitope comprises a non-nativesecondary structure of the one or plurality of self-assemblingpolypeptides.

In some embodiments, the comopsitions comprise a nucleic acid seqeunceencoding any combination of nucleic acid sequences disclosed herein orvairants thereof. In some embodiments, the comopsitions comprise a viralparticle that comprises an expressible nucleic acid seqeunce encodingany combination of nucleic acid sequences disclosed herein or variantsthereof. The component of the self-assemblying peptide can be anymonomer that, upon expression, self-assembles into a particle comprising7, 14, 27 or 60 peptides sided particle, each peptide side fused to atleast one antigen from the Coronoviridae family. In some embodiments,the composition comprises a particle comprising 7, 14, 27 or 60 peptidessided particle, each peptide side is fused to at least one antigen fromthe Coronoviridae family, wherein the antigen is positioned in anenergetically stable state as compared to the unassociated energy state.In some emboidments, the disclosure relates to a cell comprising aparticle comprising 7, 14, 27 or 60 peptides sided particle, eachpeptide side is fused to at least one antigen from the Coronoviridaefamily, wherein the antigen is positioned in an energetically stablestate as compared to the unassociated energy state. In some embodiments,the particle is able to display the at least one viral antigen in aconformationally stable state that is more immunologically active, suchactivity elicting a more therapeutically effective immune response ascompared to eh immune response elicited from administration of the DNAor RNA. In some embodiments, the energetically stable state isidentified by association of the peptide to an antibody through surfaceplasmon resonance (SPR). In some embodiments, the energetically stablestate is measured by absorbance units when either a ligand for theantigen or the antigen is immobilized to a surface, and the otherbinding partner is then passed over the surface as analyte. In someembodiments, the association can be measured through SPR on a BIACORE®system.

A detailed discussion of the technical aspects of the BIACORE®instruments and the phenomenon of SPR may be found in U.S. Pat. No.5,313,264 (the full disclosure of which is incorporated by referenceherein in its entirety). In the BIACORE® system, the SPR response valuesare expressed in resonance units (RU). One RU represents a change of0.0001° in the angle of minimum reflected light intensity. For an SPRbased sensor system like the BIACORE® system, a difference in refractiveindex between the two guiding fluids of, say, about 100 RU may beconvenient, and the fluid interface position may be determined by meansof per se conventional sensorgrams.

In some embodiments, it may be preferred to keep the total flow rateconstant when introducing the sample flow. In such a case, the flowrates of the two guiding fluids are reduced while maintaining the flowrate ratio between them. Assume, for example, that the flow rate of oneguiding fluid is 70 μl/min and the flow rate of the other guiding fluidis 30 μl/min, the total flow rate being 100 μl/min, and that a samplefluid flow of 20 μl/min is introduced between the guiding fluids. Tomaintain the total fluid flow rate at 100 μl/min, the flow rates of theguiding fluids will have to be reduced to 60 and 20 μl/min,respectively. The position of a sample fluid flow on a surface may bepresented in various ways. A non-limiting example of a experimentindicating the relative responses obtained at different detector rows asthe sample flow is guided laterally across the sensing surface of a flowcell by two guiding buffers in a BIACORE® system equipped with a ψ-cell(BIACORE® S51 is a SPR-based biosensor instrument, normally equippedwith two Y-type flow cells, each allowing a dual flow over the a sensorsurface for hydrodynamic addressing; Biacore AB, Uppsala, Sweden). Totalbuffer flow can be set to 100 μl/min, and the flow rates of the twobuffer flows can be changed in steps of 2 μl/min, starting with 2 μl/minfor one buffer and 98 μl/min for the other. Sample fluid flow can be 20μl/min all the time. Relative responses>0.1 (i.e. 10% coverage of thedetector row) are represented are measured as absorbance over time. Thisapproach thus permits convenient visual monitoring of the sample fluidflow.

In some embodiments, the stability of an antigen secondary structurewith an elevated stability as compared to a native antigen or antigennot fused to self-assemblying peptide is from about 10 to about 10,000RU more than the RU from a control as measured by SPR. In someembodiments, the stability of an antigen secondary structure with anelevated stability as compared to a native antigen or antigen not fusedto self-assemblying peptide is from about 5 to about 1,000 RU more thanthe RU from a control as measured by SPR. In some embodiments, thestability of an antigen secondary structure with an elevated stabilityas compared to a native antigen or antigen not fused to self-assemblyingpeptide is from about 100 to about 10,000 RU more than the RU from acontrol as measured by SPR. In some embodiments, the stability of anantigen secondary structure with an elevated stability as compared to anative antigen or antigen not fused to self-assemblying peptide is fromabout 100 to about 500 RU more than the RU from a control as measured bySPR. In some embodiments, the stability of an antigen secondarystructure with an elevated stability as compared to a native antigen orantigen not fused to self-assemblying peptide is from about 100 to about200 RU more than the RU from a control as measured by SPR.

5. Regulatory Sequences

In some embodiments, the expressible nucleic acid sequence can beoperably linked to one or a plurality of regulatory sequences. The term“regulatory sequence” as used herein refer to DNA sequences which arenecessary to effect expression of sequences to which they are ligated.The term “regulatory sequence” is intended to include, as a minimum, allcomponents necessary for expression and optionally additionaladvantageous components. Examples of regulatory sequences include, butnot limited to, promoters, enhancers and other expression controlelements (e.g., polyadenylation signals). Further examples of regulatorysequences are described in, for example, Goeddel, 1990, Gene ExpressionTechnology: Methods in Enzymology 185, Academic Press, San Diego, Calif.and Baron et al., 1995, Nucleic Acids Res. 23:3605-06. In someembodiments, the regulatory sequence is a promoter sequence. As usedherein, a “promoter” means a region of DNA upstream from thetranscription start and which is involved in binding RNA polymerase andother proteins to start transcription. Reference herein to a “promoter”is to be taken in its broadest context and includes the transcriptionalregulatory sequences derived from a classical eukaryotic genomic gene,including the TATA box which is required for accurate transcriptioninitiation, with or without a CCAAT box sequence and additionalregulatory elements (i.e. upstream activating sequences, enhancers andsilencers) which alter gene expression in response to developmentaland/or external stimuli, or in a tissue-specific manner. Consequently, arepressible promoter's rate of transcription decreases in response to arepressing agent. An inducible promoter's rate of transcriptionincreases in response to an inducing agent. A constitutive promoter'srate of transcription is not specifically regulated, though it can varyunder the influence of general metabolic conditions. The term “promoter”also includes the transcriptional regulatory sequences of a classicalprokaryotic gene, in which case it may include a −35 box sequence and/ora −10 box transcriptional regulatory sequences. The term “promoter” isalso used to describe a synthetic or fusion molecule, or derivativewhich confers, activates or enhances expression of a nucleic acidmolecule in a cell, tissue or organ.

6. Expressible Nucleic Acid Sequences

The expressible nucleic acid sequence comprised in the composition ofthe present disclosure can be in form of a DNA molecule, a RNA moleculeor transcript, or a DNA/RNA hybrid. In some embodiments, the expressiblenucleic acid sequence is in form of a DNA molecule. In some embodiments,the expressible nucleic acid sequence is in form of a RNA molecule ortranscript. In some embodiments, the expressible nucleic acid sequenceis in form of a DNA/RNA hybrid.

In some embodiments, the expressible nucleic acid sequence comprises afirst nucleic acid sequence encoding a self-assembling polypeptidecomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 8, or a functional fragment or variant thereof,and a second nucleic acid sequence encoding a viral antigen comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO:171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQID NO: 176 or SEQ ID NO: 177, or a functional fragment or variantthereof. In some embodiments, the expressible nucleic acid sequencecomprises a first nucleic acid sequence encoding a self-assemblingpolypeptide comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 10, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO:175, SEQ ID NO: 176 or SEQ ID NO: 177, or a functional fragment orvariant thereof. In some embodiments, the expressible nucleic acidsequence comprises a first nucleic acid sequence encoding aself-assembling polypeptide comprising at least about 60%, 65%, 70%,75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 12, or afunctional fragment or variant thereof, and a second nucleic acidsequence encoding a viral antigen comprising at least about 60%, 65%,70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60, SEQ IDNO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO: 172,SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 or SEQ IDNO: 177, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence comprises a firstnucleic acid sequence encoding a self-assembling polypeptide comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 14, or a functional fragment or variant thereof, and a secondnucleic acid sequence encoding a viral antigen comprising at least about60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60,SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO:172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 orSEQ ID NO: 177, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence comprises a firstnucleic acid sequence encoding a self-assembling polypeptide comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 16, or a functional fragment or variant thereof, and a secondnucleic acid sequence encoding a viral antigen comprising at least about60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60,SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO:172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 orSEQ ID NO: 177, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence comprises a firstnucleic acid sequence encoding a self-assembling polypeptide comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 18, or a functional fragment or variant thereof, and a secondnucleic acid sequence encoding a viral antigen comprising at least about60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60,SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO:172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 orSEQ ID NO: 177, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence comprises a firstnucleic acid sequence encoding a self-assembling polypeptide comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 20, or a functional fragment or variant thereof, and a secondnucleic acid sequence encoding a viral antigen comprising at least about60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60,SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO:172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 orSEQ ID NO: 177, or a functional fragment or variant thereof.

In some embodiments, the expressible nucleic acid sequence comprises afirst nucleic acid sequence encoding a self-assembling polypeptidecomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 8, or a functional fragment or variant thereof,and a second nucleic acid sequence encoding a viral antigen comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 175. In some embodiments, the expressible nucleic acidsequence comprises a first nucleic acid sequence encoding aself-assembling polypeptide comprising at least about 60%, 65%, 70%,75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 8, or a functionalfragment or variant thereof, and a second nucleic acid sequence encodinga viral antigen comprising at least about 60%, 65%, 70%, 75%, 80%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or100% sequence identity to SEQ ID NO: 176. In some embodiments, theexpressible nucleic acid sequence comprises a first nucleic acidsequence encoding a self-assembling polypeptide comprising at leastabout 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ IDNO: 8, or a functional fragment or variant thereof, and a second nucleicacid sequence encoding a viral antigen comprising at least about 60%,65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 177. Insome embodiments, the expressible nucleic acid sequence comprises afirst nucleic acid sequence encoding a self-assembling polypeptidecomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 12, or a functional fragment or variant thereof,and a second nucleic acid sequence encoding a viral antigen comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 175. In some embodiments, the expressible nucleic acidsequence comprises a first nucleic acid sequence encoding aself-assembling polypeptide comprising at least about 60%, 65%, 70%,75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 12, or afunctional fragment or variant thereof, and a second nucleic acidsequence encoding a viral antigen comprising at least about 60%, 65%,70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 176. In someembodiments, the expressible nucleic acid sequence comprises a firstnucleic acid sequence encoding a self-assembling polypeptide comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 12, or a functional fragment or variant thereof, and a secondnucleic acid sequence encoding a viral antigen comprising at least about60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:177.

In some embodiments, the expressible nucleic acid sequence comprises afirst nucleic acid sequence encoding a leader sequence comprising atleast about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQID NO: 1, or a functional fragment or variant thereof, and a secondnucleic acid sequence encoding a viral antigen comprising at least about60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60,SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO:172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 orSEQ ID NO: 177, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence comprises a firstnucleic acid sequence encoding a leader sequence comprising at leastabout 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ IDNO: 5, or a functional fragment or variant thereof, and a second nucleicacid sequence encoding a viral antigen comprising at least about 60%,65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60, SEQID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO: 172,SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 or SEQ IDNO: 177, or a functional fragment or variant thereof.

In some embodiments, the expressible nucleic acid sequence comprises afirst nucleic acid sequence encoding a leader sequence comprising atleast about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQID NO: 1, or a functional fragment or variant thereof, and a secondnucleic acid sequence encoding a viral antigen comprising at least about60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:171. In some embodiments, the expressible nucleic acid sequencecomprises a first nucleic acid sequence encoding a leader sequencecomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 1, or a functional fragment or variant thereof,and a second nucleic acid sequence encoding a viral antigen comprisingat least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 172. In some embodiments, the expressible nucleic acidsequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 1, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 173. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 1, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 174. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 1, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 175. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 1, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 176. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 1, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 177. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 5, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 171. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 5, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 172. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 5, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 173. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 5, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 174. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 5, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 175. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 5, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 176. In some embodiments, the expressible nucleicacid sequence comprises a first nucleic acid sequence encoding a leadersequence comprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 5, or a functional fragment or variantthereof, and a second nucleic acid sequence encoding a viral antigencomprising at least about 60%, 65%, 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 177.

Exemplary expressible nucleic acid sequences include, but not limited tothose provided in TABLE 1.

TABLE 1Exemplary Expressible Nucleic Acid Sequences (DNA and RNA) of theDisclosure and the corresponding coding polypeptide sequences (underlined amino acidresidues are glycan sites). I. CoV2 Nanoparticle ConstructsWuhanS_FP12_L9GT60_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcatgcagatctacgaaggaaaactgaccgctgagggactgaggttcggaattgtcgcaagccgcgcgaatcacgcactggtggataggctggtggaaggcgctatcgacgcaattgtccggcacggcgggagagaggaagacatcacactggtgagagtctgcggcagctgggagattcccgtggcagctggagaactggctcgaaaggaggacatcgatgccgtgatcgctattggggtcctgtgccgaggagcaactcccagcttcgactacatcgcctcagaagtgagcaaggggctggctgatctgtccctggagctgaggaaacctatcacttttggcgtgattactgccgacaccctggaacaggcaatcgaggcggccggcacctgccatggaaacaaaggctgggaagcagccctgtgcgctattgagatggcaaatctgttcaaatctctgcgaggaggctccggaggatctggagggagtggaggctcaggaggaggcgacaccatcacactgccatgccgccctgcaccacctccacattgtagctccaacatcaccggcctgattctgacaagacaggggggatatagtaacgataataccgtgattttcaggccctcaggaggggactggagggacatcgcacgatgccagattgctggaacagtggtctctactcagctgtttctgaacggcagtctggctgaggaagaggtggtcatccgatctgaagactggcgggataatgcaaagtcaatttgtgtgcagctgaacacaagcgtcgagatcaattgcactggcgcagggcactgtaacatttctcgggccaaatgggataataccctgaagcagatcgccagtaaactgagagagcagtacggcaataagacaatcatcttcaagccttctagtggaggcgacccagagttcgtgaaccatagctttaattgcgggggagagttcttttattgtgattccacacagctgttcgatagcacttggtttgattccaccggtgggagcggaagtggcggttccggatcattcattgaagaccttctctttaacaaggtgaccctcgccgatgcaggtttcattaagcaatatggtgattgcctgggagacatcgcggctcgtgatcttatttgtgcgcagaaattttaatga (SEQ ID NO: 68)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagcaugcagaucuacgaaggaaaacugaccgcugagggacugagguucggaauugucgcaagccgcgcgaaucacgcacugguggauaggcugguggaaggcgcuaucgacgcaauuguccggcacggcgggagagaggaagacaucacacuggugagagucugcggcagcugggagauucccguggcagcuggagaacuggcucgaaaggaggacaucgaugccgugaucgcuauugggguccugugccgaggagcaacucccagcuucgacuacaucgccucagaagugagcaaggggcuggcugaucugucccuggagcugaggaaaccuaucacuuuuggcgugauuacugccgacacccuggaacaggcaaucgaggcggccggcaccugccauggaaacaaaggcugggaagcagcccugugcgcuauugagauggcaaaucuguucaaaucucugcgaggaggcuccggaggaucuggagggaguggaggcucaggaggaggcgacaccaucacacugccaugccgcccugcaccaccuccacauuguagcuccaacaucaccggccugauucugacaagacaggggggauauaguaacgauaauaccgugauuuucaggcccucaggaggggacuggagggacaucgcacgaugccagauugcuggaacaguggucucuacucagcuguuucugaacggcagucuggcugaggaagagguggucauccgaucugaagacuggcgggauaaugcaaagucaauuugugugcagcugaacacaagcgucgagaucaauugcacuggcgcagggcacuguaacauuucucgggccaaaugggauaauacccugaagcagaucgccaguaaacugagagagcaguacggcaauaagacaaucaucuucaagccuucuaguggaggcgacccagaguucgugaaccauagcuuuaauugcgggggagaguucuuuuauugugauuccacacagcuguucgauagcacuugguuugauuccaccggugggagcggaaguggcgguuccggaucauucauugaagaccuucucuuuaacaaggugacccucgccgaugcagguuucauuaagcaauauggugauugccugggagacaucgcggcucgugaucuuauuugugcgcagaaauuuuaauga (SEQ ID NO: 69)MDWTWILFLVAAATRVHSMQIYEGKLTAEGLRFGIVASRANHALVDRLVEGAIDAIVRHGGREEDITLVRVCGSWEIPVAAGELARKEDIDAVIAIGVLCRGATPSFDYIASEVSKGLADLSLELRKPITFGVITADTLEQAIEAAGTCHGNKGWEAALCAIEMANLFKSLRGGSGGSGGSGGSGGGDTITLPCRPAPPPHCSSNITGLILTRQGGYSNDNTVIFRPSGGDWRDIARCQIAGTVVSTQLFLNGSLAEEEVVIRSEDWRDNAKSICVQLNTSVEINCTGA GHCNISRAKW

LKQIASKLREQYGNKTIIFKPSSGGDPEFVNHSFNCGGEFFYCDS TQLF

WF

GGSGSGGSGSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICA QKF** (SEQ ID NO: 70)WuhanS_FP_L9GT60_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcatgcagatctacgaaggaaaactgaccgctgagggactgaggttcggaattgtcgcaagccgcgcgaatcacgcactggtggataggctggtggaaggcgctatcgacgcaattgtccggcacggcgggagagaggaagacatcacactggtgagagtctgcggcagctgggagattcccgtggcagctggagaactggctcgaaaggaggacatcgatgccgtgatcgctattggggtcctgtgccgaggagcaactcccagcttcgactacatcgcctcagaagtgagcaaggggctggctgatctgtccctggagctgaggaaacctatcacttttggcgtgattactgccgacaccctggaacaggcaatcgaggcggccggcacctgccatggaaacaaaggctgggaagcagccctgtgcgctattgagatggcaaatctgttcaaatctctgcgaggaggctccggaggatctggagggagtggaggctcaggaggaggcgacaccatcacactgccatgccgccctgcaccacctccacattgtagctccaacatcaccggcctgattctgacaagacaggggggatatagtaacgataataccgtgattttcaggccctcaggaggggactggagggacatcgcacgatgccagattgctggaacagtggtctctactcagctgtttctgaacggcagtctggctgaggaagaggtggtcatccgatctgaagactggcgggataatgcaaagtcaatttgtgtgcagctgaacacaagcgtcgagatcaattgcactggcgcagggcactgtaacatttctcgggccaaatgggataataccctgaagcagatcgccagtaaactgagagagcagtacggcaataagacaatcatcttcaagccttctagtggaggcgacccagagttcgtgaaccatagctttaattgcgggggagagttcttttattgtgattccacacagctgttcgatagcacttggtttgattccaccggtgggagcggaagtggcggttccggaccttcaaagagatctttcattgaagacctgcttttcaacaaggtctaatga (SEQ ID NO: 71)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagcaugcagaucuacgaaggaaaacugaccgcugagggacugagguucggaauugucgcaagccgcgcgaaucacgcacugguggauaggcugguggaaggcgcuaucgacgcaauuguccggcacggcgggagagaggaagacaucacacuggugagagucugcggcagcugggagauucccguggcagcuggagaacuggcucgaaaggaggacaucgaugccgugaucgcuauugggguccugugccgaggagcaacucccagcuucgacuacaucgccucagaagugagcaaggggcuggcugaucugucccuggagcugaggaaaccuaucacuuuuggcgugauuacugccgacacccuggaacaggcaaucgaggcggccggcaccugccauggaaacaaaggcugggaagcagcccugugcgcuauugagauggcaaaucuguucaaaucucugcgaggaggcuccggaggaucuggagggaguggaggcucaggaggaggcgacaccaucacacugccaugccgcccugcaccaccuccacauuguagcuccaacaucaccggccugauucugacaagacaggggggauauaguaacgauaauaccgugauuuucaggcccucaggaggggacuggagggacaucgcacgaugccagauugcuggaacaguggucucuacucagcuguuucugaacggcagucuggcugaggaagagguggucauccgaucugaagacuggcgggauaaugcaaagucaauuugugugcagcugaacacaagcgucgagaucaauugcacuggcgcagggcacuguaacauuucucgggccaaaugggauaauacccugaagcagaucgccaguaaacugagagagcaguacggcaauaagacaaucaucuucaagccuucuaguggaggcgacccagaguucgugaaccauagcuuuaauugcgggggagaguucuuuuauugugauuccacacagcuguucgauagcacuugguuugauuccaccggugggagcggaaguggcgguuccggaccuucaaagagaucuuucauugaagaccugcuuuucaacaaggucuaauga (SEQ ID NO: 72)MDWTWILFLVAAATRVHSMQIYEGKLTAEGLRFGIVASRANHALVDRLVEGAIDAIVRHGGREEDITLVRVCGSWEIPVAAGELARKEDIDAVIAIGVLCRGATPSFDYIASEVSKGLADLSLELRKPITFGVITADTLEQAIEAAGTCHGNKGWEAALCAIEMANLFKSLRGGSGGSGGSGGSGGGDTITLPCRPAPPPHCSSNITGLILTRQGGYSNDNTVIFRPSGGDWRDIARCQIAGTVVSTQLFLNGSLAEEEVVIRSEDWRDNAKSICVQLNTSVEINCTGA GHCNISRAKW

LKQIASKLREQYGNKTIIFKPSSGGDPEFVNHSFNCGGEFFYCDS TQLF

WF

GGSGSGGSGPSKRSFIEDLLFNKV** (SEQ ID NO: 73) WuhanS_RBD_gmax_180_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcctctcaattgccccaacgttgatcaaccgggacaagccatacacgaaagaggaacttatggagatattgcggttggccattatagctgaactcgatgcaattaatctctatgaacaaatggcccgctatagcgaagacgaaaatgtgagaaagatcttgttggacgtcgctagggaagagaaagcacacgtaggagagttcatggctttgttgcttaacctcgaccctgagcaagtcacagagctgaagggcgggttcgaggaagttaaagaattgaccggtatagaagctcacattaatgacaacaagaaagaggaaagtaatgtagagtatttcgagaagctcagatctgccttgttggatggagtcaacaagggtcgcagcttgctcaaacatctgcccgttacaagaatagaagggcagtcttttcgagtagacatcatcaaatttgaggacggcgtccgagtggttaaacaagagtataagcctataccccttcttaagaagaagttctacgtcggcattcgagaactgaatgacgggacatatgatgtcagcattgctactaaagccggtgagctgctggttaaagacgaagaaagtcttgtgatccgggaaattttgtcaacggaaggcatcaagaaaatgaaattgtcatcctgggacaatccagaagaagccctgaatgatttgatgaatgcgctccaagaagctagcaatgctagtgctggccccttcggccttattatcaatccaaagcggtacgccaaactgctgaagatctatgaaaagtcaggtaagatgctcgtagaagtactcaaggaaatcttccggggtggaataatcgtaactcttaatatcgacgaaaacaaagtgattatcttcgctaatacgcccgccgttctggacgtggtggtgggtcaagacgttacgctccaggagcttggtccggaaggggatgatgtcgcattcctggtcagtgaagccattggtataagaatcaagaacccggaagctatagttgttctcgaaggcgggtctggtgggagcggtggtagtggtggttctggtggtggtgggtcaggtggcggctcaggcggcggcaatctgtgccctttcggtgaggtctttaatgcaacaagatttgcaagtgtttacgcctggaaccgtaagcgcattagcaactgcactgccgattactctgtgctgtacaacagcacaagcttttccacatttaaatgttacggggtttcccctaccaacctcagcgacctctgctttactaatgtttacgcagattccttcgttatccgaggcgatgaagtccggcagatagctcccggacagaccggcaaaatcgctgactacaactataaactgccgaacgacagcacagggtgtgtaattgcttggaacagcaataacctcgattcaaaggttggcggaaattacaattatctttaccgtctgttccggaaaagcaatctgaaaccctttgagagagacatcagcacggaaatttatcaagccggttcaacaccatgtaacggagttgaaggctttaattgctattttcccctgcaatcttacggatttcaacctacgaacggggtcggttaccaaccttaccgggtggtcgtgctgagcttcgaattgcttcatgccccagccaccgtgtgtgggccataatga (SEQ ID NO: 74)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagccucucaauugccccaacguugaucaaccgggacaagccauacacgaaagaggaacuuauggagauauugcgguuggccauuauagcugaacucgaugcaauuaaucucuaugaacaaauggcccgcuauagcgaagacgaaaaugugagaaagaucuuguuggacgucgcuagggaagagaaagcacacguaggagaguucauggcuuuguugcuuaaccucgacccugagcaagucacagagcugaagggcggguucgaggaaguuaaagaauugaccgguauagaagcucacauuaaugacaacaagaaagaggaaaguaauguagaguauuucgagaagcucagaucugccuuguuggauggagucaacaagggucgcagcuugcucaaacaucugcccguuacaagaauagaagggcagucuuuucgaguagacaucaucaaauuugaggacggcguccgagugguuaaacaagaguauaagccuauaccccuucuuaagaagaaguucuacgucggcauucgagaacugaaugacgggacauaugaugucagcauugcuacuaaagccggugagcugcugguuaaagacgaagaaagucuugugauccgggaaauuuugucaacggaaggcaucaagaaaaugaaauugucauccugggacaauccagaagaagcccugaaugauuugaugaaugcgcuccaagaagcuagcaaugcuagugcuggccccuucggccuuauuaucaauccaaagcgguacgccaaacugcugaagaucuaugaaaagucagguaagaugcucguagaaguacucaaggaaaucuuccgggguggaauaaucguaacucuuaauaucgacgaaaacaaagugauuaucuucgcuaauacgcccgccguucuggacgugguggugggucaagacguuacgcuccaggagcuugguccggaaggggaugaugucgcauuccuggucagugaagccauugguauaagaaucaagaacccggaagcuauaguuguucucgaaggcgggucuggugggagcggugguaguggugguucuggugguggugggucagguggcggcucaggcggcggcaaucugugcccuuucggugaggucuuuaaugcaacaagauuugcaaguguuuacgccuggaaccguaagcgcauuagcaacugcacugccgauuacucugugcuguacaacagcacaagcuuuuccacauuuaaauguuacgggguuuccccuaccaaccucagcgaccucugcuuuacuaauguuuacgcagauuccuucguuauccgaggcgaugaaguccggcagauagcucccggacagaccggcaaaaucgcugacuacaacuauaaacugccgaacgacagcacaggguguguaauugcuuggaacagcaauaaccucgauucaaagguuggcggaaauuacaauuaucuuuaccgucuguuccggaaaagcaaucugaaacccuuugagagagacaucagcacggaaauuuaucaagccgguucaacaccauguaacggaguugaaggcuuuaauugcuauuuuccccugcaaucuuacggauuucaaccuacgaacggggucgguuaccaaccuuaccggguggucgugcugagcuucgaauugcuucaugccccagccaccgugugugggccauaauga (SEQ ID NO: 75)MDWTWILFLVAAATRVHSLSIAPTLINRDKPYTKEELMEILRLAIIAELDAINLYEQMARYSEDENVRKILLDVAREEKAHVGEFMALLLNLDPEQVTELKGGFEEVKELTGIEAHINDNKKEESNVEYFEKLRSALLDGVNKGRSLLKHLPVTRIEGQSFRVDIIKFEDGVRVVKQEYKPIPLLKKKFYVGIRELNDGTYDVSIATKAGELLVKDEESLVIREILSTEGIKKMKLSSWDNPEEALNDLMNALQEASNASAGPFGLIINPKRYAKLLKIYEKSGKMLVEVLKEIFRGGIIVTLNIDENKVIIFANTPAVLDVVVGQDVTLQELGPEGDDVAFLVSEAIGIRIKNPEAIVVLEGGSGGSGGSGGSGGGGSGGGSGGGNLCPFGEVFNATRFASVYAWNRKRISNCTADYSVLYNSTSFSTFKCYGVSPTNLSDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPNDSTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGP** (SEQ ID NO: 76) WuhanS_RBD_gmax_LS_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcatgcagatctacgaaggaaaactgaccgctgagggactgaggttcggaattgtcgcaagccgcgcgaatcacgcactggtggataggctggtggaaggcgctatcgacgcaattgtccggcacggcgggagagaggaagacatcacactggtgagagtctgcggcagctgggagattcccgtggcagctggagaactggctcgaaaggaggacatcgatgccgtgatcgctattggggtcctgtgccgaggagcaactcccagcttcgactacatcgcctcagaagtgagcaaggggctggctgatctgtccctggagctgaggaaacctatcacttttggcgtgattactgccgacaccctggaacaggcaatcgaggcggccggcacctgccatggaaacaaaggctgggaagcagccctgtgcgctattgagatggcaaatctgttcaaatctctgcgaggaggctccggaggatctggagggagtggaggctcaggaggaggcaatctgtgccctttcggtgaggtctttaatgcaacaagatttgcaagtgtttacgcctggaaccgtaagcgcattagcaactgcactgccgattactctgtgctgtacaacagcacaagcttttccacatttaaatgttacggggtttcccctaccaacctcagcgacctctgctttactaatgtttacgcagattccttcgttatccgaggcgatgaagtccggcagatagctcccggacagaccggcaaaatcgctgactacaactataaactgccgaacgacagcacagggtgtgtaattgcttggaacagcaataacctcgattcaaaggttggcggaaattacaattatctttaccgtctgttccggaaaagcaatctgaaaccctttgagagagacatcagcacggaaatttatcaagccggttcaacaccatgtaacggagttgaaggctttaattgctattttcccctgcaatcttacggatttcaacctacgaacggggtcggttaccaaccttaccgggtggtcgtgctgagcttcgaattgcttcatgccccagccaccgtgtgtgggccataatga (SEQ ID NO: 77)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagcaugcagaucuacgaaggaaaacugaccgcugagggacugagguucggaauugucgcaagccgcgcgaaucacgcacugguggauaggcugguggaaggcgcuaucgacgcaauuguccggcacggcgggagagaggaagacaucacacuggugagagucugcggcagcugggagauucccguggcagcuggagaacuggcucgaaaggaggacaucgaugccgugaucgcuauugggguccugugccgaggagcaacucccagcuucgacuacaucgccucagaagugagcaaggggcuggcugaucugucccuggagcugaggaaaccuaucacuuuuggcgugauuacugccgacacccuggaacaggcaaucgaggcggccggcaccugccauggaaacaaaggcugggaagcagcccugugcgcuauugagauggcaaaucuguucaaaucucugcgaggaggcuccggaggaucuggagggaguggaggcucaggaggaggcaaucugugcccuuucggugaggucuuuaaugcaacaagauuugcaaguguuuacgccuggaaccguaagcgcauuagcaacugcacugccgauuacucugugcuguacaacagcacaagcuuuuccacauuuaaauguuacgggguuuccccuaccaaccucagcgaccucugcuuuacuaauguuuacgcagauuccuucguuauccgaggcgaugaaguccggcagauagcucccggacagaccggcaaaaucgcugacuacaacuauaaacugccgaacgacagcacaggguguguaauugcuuggaacagcaauaaccucgauucaaagguuggcggaaauuacaauuaucuuuaccgucuguuccggaaaagcaaucugaaacccuuugagagagacaucagcacggaaauuuaucaagccgguucaacaccauguaacggaguugaaggcuuuaauugcuauuuuccccugcaaucuuacggauuucaaccuacgaacggggucgguuaccaaccuuaccggguggucgugcugagcuucgaauugcuucaugccccagccaccgugugugggccauaauga (SEQ ID NO: 78)MDWTWILFLVAAATRVHSMQIYEGKLTAEGLRFGIVASRANHALVDRLVEGAIDAIVRHGGREEDITLVRVCGSWEIPVAAGELARKEDIDAVIAIGVLCRGATPSFDYIASEVSKGLADLSLELRKPITFGVITADTLEQAIEAAGTCHGNKGWEAALCAIEMANLFKSLRGGSGGSGGSGGSGGGNLCPFGEVFNATRFASVYAWNRKRISNCTADYSVLYNSTSFSTFKCYGVSPTNLSDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPNDSTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGP** (SEQ ID NO: 79)CoV2-RBD_7 mer_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcaatctctgtccattcggagaggttttcaacgcgacgagattcgcctcagtttatgcctggaaccgtaaacggatatcaaactgcgtggctgactactctgttttatacaactccgcctctttcagtaccttcaagtgttacggtgtcagccctaccaaattgaatgatctctgctttacaaatgtttacgcagattcttttgtcataaggggcgatgaggttcggcaaatcgcccccgggcagacaggcaaaattgcggactataattataagttgccagacgatttcacgggctgcgtcatcgcctggaacagtaataatctcgattcaaaagtgggtgggaactacaattatctctacaggttattccggaagtcaaatctgaagcccttcgaacgcgacatcagtacggagatttaccaggctggaagcactccgtgcaacggggtggaggggttcaactgttattttcctctgcagtcttatgggtttcagcccactaatggtgtgggataccagccgtacagagtcgtggtgctgtccttcgaacttctccacgctcccgccaccgtctgtggtcccgggggatctggcggatcagggggtagtggaggtagcggcggcgggaagaaacagggagacgctgacgtctgtggggaagtggcttacatccagagcgtggtgtctgattgccatgtaccaaccgcggagctcaggactcttttagagattcggaaactgtttctggagatccaaaagctgaaggtcgaactccagggcctgtcaaaagaatgataa (SEQ ID NO: 80)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagcaaucucuguccauucggagagguuuucaacgcgacgagauucgccucaguuuaugccuggaaccguaaacggauaucaaacugcguggcugacuacucuguuuuauacaacuccgccucuuucaguaccuucaaguguuacggugucagcccuaccaaauugaaugaucucugcuuuacaaauguuuacgcagauucuuuugucauaaggggcgaugagguucggcaaaucgcccccgggcagacaggcaaaauugcggacuauaauuauaaguugccagacgauuucacgggcugcgucaucgccuggaacaguaauaaucucgauucaaaagugggugggaacuacaauuaucucuacagguuauuccggaagucaaaucugaagcccuucgaacgcgacaucaguacggagauuuaccaggcuggaagcacuccgugcaacgggguggagggguucaacuguuauuuuccucugcagucuuauggguuucagcccacuaauggugugggauaccagccguacagagucguggugcuguccuucgaacuucuccacgcucccgccaccgucuguggucccgggggaucuggcggaucaggggguaguggagguagcggcggcgggaagaaacagggagacgcugacgucuguggggaaguggcuuacauccagagcguggugucugauugccauguaccaaccgcggagcucaggacucuuuuagagauucggaaacuguuucuggagauccaaaagcugaaggucgaacuccagggccugucaaaagaaugauaa(SEQ ID NO: 81)NLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPGGSGGSGGSGGSGGGKKQGDADVCGEVAYIQSVVSDCHVPTAELRTLLEIRKLFLEIQKLKVELQGLSKE** (SEQ ID NO: 82)CoV2-RBD_gmax_14 mer_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcaatctgtgtcccttcggggaggttttcaatgctaccagatttgccagtgtgtatgcttggaatcggaagagaatctccaattgcacagcagattattcagttctctacaactctacatcttttagtacctttaagtgttacggggtgagtcccactaacctttcagatttatgtttcaccaatgtctacgctgactccttcgtgatccggggggatgaggtgagacagattgcacctggacaaactggcaaaatcgccgactacaattacaaacttccaaacgactctacagggtgtgtaatcgcttggaacagcaataatctggatagcaaagtaggcggcaattataattacctctacagactgtttaggaagtccaacctgaaaccatttgagagggacatcagcactgaaatctaccaggcggggagcaccccttgtaatggagtcgagggtttcaactgttacttcccactgcagagctacgggttccagcctaccaatggtgtcggttaccagccctatcgagttgtggtgttgtcattcgaactgttacatgcacctgcaacggtctgtggacccgggggttcagggggtagtggggggtccggtgggagcggtgggggcaagaaacaggggatgaatccgctcatcgccgccgcctctgtgatagctgctggcctggccgtgggcctggcatcaatcgggcccggggtgggccaaggcaccgccgccggccaggccgtcgagggtattgcaaggcagccggaggcagaaggcaaaattagagggaccctgttgttgtctttagcgttcatggaagccctcactatttacggactggttgtggccttagcccttctgtttgccaatcctttcgtgtaatga (SEQ ID NO: 83)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagcaaucugugucccuucggggagguuuucaaugcuaccagauuugccaguguguaugcuuggaaucggaagagaaucuccaauugcacagcagauuauucaguucucuacaacucuacaucuuuuaguaccuuuaaguguuacggggugagucccacuaaccuuucagauuuauguuucaccaaugucuacgcugacuccuucgugauccggggggaugaggugagacagauugcaccuggacaaacuggcaaaaucgccgacuacaauuacaaacuuccaaacgacucuacaggguguguaaucgcuuggaacagcaauaaucuggauagcaaaguaggcggcaauuauaauuaccucuacagacuguuuaggaaguccaaccugaaaccauuugagagggacaucagcacugaaaucuaccaggcggggagcaccccuuguaauggagucgaggguuucaacuguuacuucccacugcagagcuacggguuccagccuaccaauggugucgguuaccagcccuaucgaguugugguguugucauucgaacuguuacaugcaccugcaacggucuguggacccggggguucaggggguagugggggguccggugggagcggugggggcaagaaacaggggaugaauccgcucaucgccgccgccucugugauagcugcuggccuggccgugggccuggcaucaaucgggcccggggugggccaaggcaccgccgccggccaggccgucgaggguauugcaaggcagccggaggcagaaggcaaaauuagagggacccuguuguugucuuuagcguucauggaagcccucacuauuuacggacugguuguggccuuagcccuucuguuugccaauccuuucguguaauga (SEQ ID NO: 84)NLCPFGEVFNATRFASVYAWNRKRISNCTADYSVLYNSTSFSTFKCYGVSPTNLSDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPNDSTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPGGSGGSGGSGGSGGGKKQGMNPLIAAASVIAAGLAVGLASIGPGVGQGTAAGQAVEGIARQPEAEGKIRGTLLLSLAFMEALTIYGLVVALALLFANPFV** (SEQ ID NO: 85) CoV2-RBD_gmax_24 mer_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcaacctgtgccccttcggggaagtttttaatgccactcggtttgcctccgtttacgcctggaacaggaagcgcatttccaactgcaccgccgactatagcgtcctttataacagcacatccttttcaactttcaagtgttacggggtttcccctacaaatctctctgacctgtgttttacaaatgtgtatgcagactctttcgtgattaggggagatgaggtgcgccagatcgcccctggacagaccggaaaaatcgccgattataattataagcttcccaacgactccacaggctgtgtaattgcctggaattctaataacctggactctaaagtgggcggtaactacaattatctgtatagactcttcagaaagtctaacctcaaaccatttgaacgggacatctcaaccgagatctaccaagccgggtccaccccctgtaacggcgtggaaggcttcaactgttatttccccctccagtcctatggcttccaacccacaaatggagtcggctaccagccttacagggtggttgtgctgtcatttgagctcctccacgctcctgccactgtatgtgggccaggcgggtccggaggttcaggcggtagcggcggctcaggtggaggaggactgtctaaagatattataaaactgctgaacgaacaagtgaacaaggagatgcagagcagcaacctttacatgtctatgagcagttggtgttacactcactctctcgacggcgccggcctgttcctgtttgatcacgccgcggaggagtatgaacatgctaaaaagcttatcatcttcctcaacgaaaataacgtgccagtgcagttgacctctatttccgctcccgaacataagttcgaaggcctcacacagatctttcagaaggcttacgagcatgaacaacacatttcagagagcatcaacaacatcgtggaccatgcgatcaagtctaaggaccacgcgacttttaacttcctccagtggtatgtcgccgaacagcatgaggaggaagtgttgttcaaagacatcctggacaagattgaacttattggcaacgaaaaccacggcctctacctggccgatcagtacgtgaaaggtatcgcgaagtcacgaaagagttaatga (SEQ ID NO: 86)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagcaaccugugccccuucggggaaguuuuuaaugccacucgguuugccuccguuuacgccuggaacaggaagcgcauuuccaacugcaccgccgacuauagcguccuuuauaacagcacauccuuuucaacuuucaaguguuacgggguuuccccuacaaaucucucugaccuguguuuuacaaauguguaugcagacucuuucgugauuaggggagaugaggugcgccagaucgccccuggacagaccggaaaaaucgccgauuauaauuauaagcuucccaacgacuccacaggcuguguaauugccuggaauucuaauaaccuggacucuaaagugggcgguaacuacaauuaucuguauagacucuucagaaagucuaaccucaaaccauuugaacgggacaucucaaccgagaucuaccaagccggguccacccccuguaacggcguggaaggcuucaacuguuauuucccccuccaguccuauggcuuccaacccacaaauggagucggcuaccagccuuacagggugguugugcugucauuugagcuccuccacgcuccugccacuguaugugggccaggcggguccggagguucaggcgguagcggcggcucagguggaggaggacugucuaaagauauuauaaaacugcugaacgaacaagugaacaaggagaugcagagcagcaaccuuuacaugucuaugagcaguugguguuacacucacucucucgacggcgccggccuguuccuguuugaucacgccgcggaggaguaugaacaugcuaaaaagcuuaucaucuuccucaacgaaaauaacgugccagugcaguugaccucuauuuccgcucccgaacauaaguucgaaggccucacacagaucuuucagaaggcuuacgagcaugaacaacacauuucagagagcaucaacaacaucguggaccaugcgaucaagucuaaggaccacgcgacuuuuaacuuccuccagugguaugucgccgaacagcaugaggaggaaguguuguucaaagacauccuggacaagauugaacuuauuggcaacgaaaaccacggccucuaccuggccgaucaguacgugaaagguaucgcgaagucacgaaagaguuaauga (SEQ ID NO: 87)NLCPFGEVFNATRFASVYAWNRKRISNCTADYSVLYNSTSFSTFKCYGVSPTNLSDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPNDSTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPGGSGGSGGSGGSGGGGLSKDIIKLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEEYEHAKKLIIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYEHEQHISESINNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIELIGNENHGLYLADQYVKGIAKSRKS** (SEQ ID NO: 88)CoV2-RBD_gmax_7 mer_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcaatctttgtccattcggggaagtgtttaacgccactaggttcgctagtgtgtacgcctggaatcggaagcggatttcaaattgtaccgccgattattctgtcctttacaacagtaccagcttttccacttttaaatgctacggagtatctcctacaaacttgagtgacctgtgttttacgaacgtctacgctgactctttcgttattaggggagacgaagttagacaaatcgctccaggccagactggcaaaatagccgactataactataaactcccaaacgattccacaggctgcgttattgcctggaacagcaataacctggactctaaagtcggaggtaactataactacttgtacaggctcttccgcaagagcaaccttaagccatttgagcgagatatctccaccgagatttatcaggcagggagcaccccatgcaacggagtggaggggtttaattgctattttccactgcagtcctatggctttcaaccaacaaacggagtaggctaccaaccgtatcgcgttgtcgtcctgagtttcgaactgttgcacgcccctgcgaccgtatgtggccccggcggctcaggggggagtggtgggagcgggggctctgggggggggaaaaaacagggggacgccgatgtttgcggcgaggtggcctatatacagtcagtggtctccgactgtcatgtaccaactgccgaactcaggactcttctggagataaggaagttgttcctggagatacagaagctcaaggtcgagttacagggtctctcaaaggaatgatga (SEQ ID NO: 89)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagcaaucuuuguccauucggggaaguguuuaacgccacuagguucgcuaguguguacgccuggaaucggaagcggauuucaaauuguaccgccgauuauucuguccuuuacaacaguaccagcuuuuccacuuuuaaaugcuacggaguaucuccuacaaacuugagugaccuguguuuuacgaacgucuacgcugacucuuucguuauuaggggagacgaaguuagacaaaucgcuccaggccagacuggcaaaauagccgacuauaacuauaaacucccaaacgauuccacaggcugcguuauugccuggaacagcaauaaccuggacucuaaagucggagguaacuauaacuacuuguacaggcucuuccgcaagagcaaccuuaagccauuugagcgagauaucuccaccgagauuuaucaggcagggagcaccccaugcaacggaguggagggguuuaauugcuauuuuccacugcaguccuauggcuuucaaccaacaaacggaguaggcuaccaaccguaucgcguugucguccugaguuucgaacuguugcacgccccugcgaccguauguggccccggcggcucaggggggaguggugggagcgggggcucugggggggggaaaaaacagggggacgccgauguuugcggcgagguggccuauauacagucaguggucuccgacugucauguaccaacugccgaacucaggacucuucuggagauaaggaaguuguuccuggagauacagaagcucaaggucgaguuacagggucucucaaaggaaugauga (SEQID NO: 90) NLCPFGEVFNATRFASVYAWNRKRISNCTADYSVLYNSTSFSTFKCYGVSPTNLSDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPNDSTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPGGSGGSGGSGGSGGGKKQGDADVCGEVAYIQSVVSDCHVPTAELRTLLEIRKLFLEIQKLKVELQGLSKE** (SEQ ID NO: 91)II. CoV2 Trimer Constructs WuhanS_SolTri_FS1_pVaxggatccgccaccatggactggacctggattctgttcctggtggccgccgccacaagggtgcacagcatgtttgttttcctcgtcctcttgcccctcgtctctagtcaatgtgttaatttgaccacacgaacccaactgccacctgcctacaccaacagttttaccagaggagtttattaccccgacaaagtattcaggtcatcagtgctgcatagtacccaagacttgtttctccccttctttagtaacgttacatggttccacgccattcacgtgagtgggacaaatggaacaaaacgcttcgacaaccctgtgctccccttcaacgatggtgtatactttgctagtaccgagaagagcggaattatccgcgggtggatctttggaacaacactggacagcaaaacccaaagcctgcttatcgttaacaatgctactaacgttgtgatcaaagtgtgtgaattccaattttgtaatgatccgtttctcggagtttactaccacaagaacaacaaaagttggatggaaagcgaattccgggtgtactcctcagcaaataattgtacctttgagtacgtgagtcaaccctttctcatggacctggaaggaaaacaaggcaatttcaagaacctgcgggagtttgtgttcaagaatattgatggctattttaaaatttattctaagcatactccaatcaacctggtaagggacctgccccaaggcttttcagccctcgaaccgcttgtagatttgcctatcgggataaacattacgcgatttcaaacgctgttggcgctccaccggagctacttgactcctggcgatagcagctccggttggaccgctggagcggccgcttattacgtcggctatctgcaacccaggacgttcctgctcaagtataatgagaacgggacgattacagatgcagtggattgtgcgcttgatcctctctctgaaaccaagtgcactctcaagtctttcacggtggagaaaggcatttatcaaactagtaactttcgagtacagcctactgagagtatcgttaggttcccaaacattacgaacctctgtccctttggagaagtattcaatgctactcgctttgcaagcgtttatgcctggaatcgcaaacgcatcagcaattgcgtcgccgattattctgtcctttataatagcgcatcattttcaacatttaagtgttatggggtgagtccgactaagctcaatgatttgtgcttcacaaacgtctacgcggacagctttgtgataaggggcgacgaagttcgccaaatcgctcccggccaaactgggaaaatcgcggattacaactataaattgcccgatgacttcaccggctgtgtcattgcctggaactctaataacctcgatagcaaggtgggcgggaactataattatttgtaccgcctgtttcgaaagtccaatctcaaaccctttgagcgggacatttccactgagatctatcaggcagggagtacaccttgtaacggcgtggaaggctttaactgttattttcccctgcaaagttacggttttcaacctaccaacggagttggctatcaaccttatcgagtcgtcgtgctgagttttgagttgctgcatgccccagccaccgtctgtggacctaagaaatccaccaacctcgtgaagaacaagtgcgtcaattttaattttaacggcctgactgggaccggtgtcctcaccgaatctaataagaagttcctgccatttcaacaattcggacgggacatcgctggaacgacagatgctgtccgtgatcctcagacactggagattctggacatcactccttgcagctttggcggagtctctgttattactcccggaactaacacttctaaccaagttgctgtcctctatcaggacgtgaactgcactgaagtgcccgtggcaatccatgcaggccaactgacccccacttggagagtctacagcacggggagcaatgtcttccaaacaagggccggatgccttattggagcggagcacgttaataactcatacgagtgtgatataccaattggagcaggaatttgtgcttcctaccagacccaaactaacagtcccaggcgggctaggagtgtcgctagccagagcatcatcgcgtacacaatgtctctcggcgcagaaaactcagtcgcctatagcaacaactcaattgccattcccaccaacttcacaatttccgtaaccactgaaattctgcctgttagcatgacaaagacatcagttgattgtacaatgtacatatgtggagacagcaccgaatgcagcaaccttttgcttcaatatggctccttttgtacccaactcaacagggcactcactgggatagcagtcgaacaagataagaacacccaagaggtgtttgcacaagtcaaacaaatctataaaacgccgcccataaaagactttggcggattcaatttcagccagatcttgcctgacccatccaagccttcaaagaggagctttattgaggatcttctcttcaataaagtgacactggcggacgccggttttatcaagcaatatggtgattgtctcggtgacatagcagctagagatctgatttgcgctcagaaatttaatggccttactgtgcttcccccactgctgaccgatgaaatgattgcacaatatacaagcgcccttttggccgggactattacttccgggtggaccttcggcgccggcgccgctctgcaaattcctttcgcaatgcagatggcctaccggttcaatggcataggtgtcactcagaacgttctttatgagaatcagaaactcatcgcgaaccagtttaattcagcgatcggcaagattcaggactccttgtcctcaactgcgtcagctttgggaaaacttcaagacgtcgtgaaccagaatgctcaggcgctcaataccctggtgaaacaacttagcagtaactttggggctatttctagcggtccaaacgatatactgtcccgactcccgaaagtcgaggccgaagtccaaattgatcgtcttattacagggagactccaatctcttcaaacatatgtcactcaacagctcattagggctgcggagatccgggcttccgcaaatcttgccgcgacaaagatgagtgaatgcgtcttgggacaatctaagagggtggacttttgtggaaaaggttaccatctcatgtccttccctcagtcagcgccccacggagtcgttttcctgcacgtaacgtatgtcccggctcaagagaagaacttcactactgcaccagcgatttgccatgacggtaaagcccattttccccgcgagggcgtatttgtgtccaacggtacccactggttcgtaacccaacggaatttctatgagccccaaatcattacaacagataatacagatgtttccgggaattgcgacgttgttattggcatcgttaacaacaccgtttacgatcccttgcaaccggaactggactcctttaaagaagaactcgacaagtattttaagaaccacacatcaccagatgtcgatcttggcgacatttccggcattaacgcttcagttgtaaatattcagaaagagatagatcgcctgaatgaggtggctaagaacctgaacgaatctctcattgatctccaagagctgggaaagtacgaacaatacatcaaatggccttctgggcgtcgccgaagacgacgagggtccggcggctcagggagcggctatatccctgaggcgcctcgggacggacaagcttatgtgaggaaagatggagaatgggtattgctgtcaaccttcctgggataatga (SEQ ID NO: 92)ggauccgccaccauggacuggaccuggauucuguuccugguggccgccgccacaagggugcacagcauguuuguuuuccucguccucuugccccucgucucuagucaauguguuaauuugaccacacgaacccaacugccaccugccuacaccaacaguuuuaccagaggaguuuauuaccccgacaaaguauucaggucaucagugcugcauaguacccaagacuuguuucuccccuucuuuaguaacguuacaugguuccacgccauucacgugagugggacaaauggaacaaaacgcuucgacaacccugugcuccccuucaacgaugguguauacuuugcuaguaccgagaagagcggaauuauccgcggguggaucuuuggaacaacacuggacagcaaaacccaaagccugcuuaucguuaacaaugcuacuaacguugugaucaaagugugugaauuccaauuuuguaaugauccguuucucggaguuuacuaccacaagaacaacaaaaguuggauggaaagcgaauuccggguguacuccucagcaaauaauuguaccuuugaguacgugagucaacccuuucucauggaccuggaaggaaaacaaggcaauuucaagaaccugcgggaguuuguguucaagaauauugauggcuauuuuaaaauuuauucuaagcauacuccaaucaaccugguaagggaccugccccaaggcuuuucagcccucgaaccgcuuguagauuugccuaucgggauaaacauuacgcgauuucaaacgcuguuggcgcuccaccggagcuacuugacuccuggcgauagcagcuccgguuggaccgcuggagcggccgcuuauuacgucggcuaucugcaacccaggacguuccugcucaaguauaaugagaacgggacgauuacagaugcaguggauugugcgcuugauccucucucugaaaccaagugcacucucaagucuuucacgguggagaaaggcauuuaucaaacuaguaacuuucgaguacagccuacugagaguaucguuagguucccaaacauuacgaaccucugucccuuuggagaaguauucaaugcuacucgcuuugcaagcguuuaugccuggaaucgcaaacgcaucagcaauugcgucgccgauuauucuguccuuuauaauagcgcaucauuuucaacauuuaaguguuauggggugaguccgacuaagcucaaugauuugugcuucacaaacgucuacgcggacagcuuugugauaaggggcgacgaaguucgccaaaucgcucccggccaaacugggaaaaucgcggauuacaacuauaaauugcccgaugacuucaccggcugugucauugccuggaacucuaauaaccucgauagcaaggugggcgggaacuauaauuauuuguaccgccuguuucgaaaguccaaucucaaacccuuugagcgggacauuuccacugagaucuaucaggcagggaguacaccuuguaacggcguggaaggcuuuaacuguuauuuuccccugcaaaguuacgguuuucaaccuaccaacggaguuggcuaucaaccuuaucgagucgucgugcugaguuuugaguugcugcaugccccagccaccgucuguggaccuaagaaauccaccaaccucgugaagaacaagugcgucaauuuuaauuuuaacggccugacugggaccgguguccucaccgaaucuaauaagaaguuccugccauuucaacaauucggacgggacaucgcuggaacgacagaugcuguccgugauccucagacacuggagauucuggacaucacuccuugcagcuuuggcggagucucuguuauuacucccggaacuaacacuucuaaccaaguugcuguccucuaucaggacgugaacugcacugaagugcccguggcaauccaugcaggccaacugacccccacuuggagagucuacagcacggggagcaaugucuuccaaacaagggccggaugccuuauuggagcggagcacguuaauaacucauacgagugugauauaccaauuggagcaggaauuugugcuuccuaccagacccaaacuaacagucccaggcgggcuaggagugucgcuagccagagcaucaucgcguacacaaugucucucggcgcagaaaacucagucgccuauagcaacaacucaauugccauucccaccaacuucacaauuuccguaaccacugaaauucugccuguuagcaugacaaagacaucaguugauuguacaauguacauauguggagacagcaccgaaugcagcaaccuuuugcuucaauauggcuccuuuuguacccaacucaacagggcacucacugggauagcagucgaacaagauaagaacacccaagagguguuugcacaagucaaacaaaucuauaaaacgccgcccauaaaagacuuuggcggauucaauuucagccagaucuugccugacccauccaagccuucaaagaggagcuuuauugaggaucuucucuucaauaaagugacacuggcggacgccgguuuuaucaagcaauauggugauugucucggugacauagcagcuagagaucugauuugcgcucagaaauuuaauggccuuacugugcuucccccacugcugaccgaugaaaugauugcacaauauacaagcgcccuuuuggccgggacuauuacuuccggguggaccuucggcgccggcgccgcucugcaaauuccuuucgcaaugcagauggccuaccgguucaauggcauaggugucacucagaacguucuuuaugagaaucagaaacucaucgcgaaccaguuuaauucagcgaucggcaagauucaggacuccuuguccucaacugcgucagcuuugggaaaacuucaagacgucgugaaccagaaugcucaggcgcucaauacccuggugaaacaacuuagcaguaacuuuggggcuauuucuagcgguccaaacgauauacugucccgacucccgaaagucgaggccgaaguccaaauugaucgucuuauuacagggagacuccaaucucuucaaacauaugucacucaacagcucauuagggcugcggagauccgggcuuccgcaaaucuugccgcgacaaagaugagugaaugcgucuugggacaaucuaagaggguggacuuuuguggaaaagguuaccaucucauguccuucccucagucagcgccccacggagucguuuuccugcacguaacguaugucccggcucaagagaagaacuucacuacugcaccagcgauuugccaugacgguaaagcccauuuuccccgcgagggcguauuuguguccaacgguacccacugguucguaacccaacggaauuucuaugagccccaaaucauuacaacagauaauacagauguuuccgggaauugcgacguuguuauuggcaucguuaacaacaccguuuacgaucccuugcaaccggaacuggacuccuuuaaagaagaacucgacaaguauuuuaagaaccacacaucaccagaugucgaucuuggcgacauuuccggcauuaacgcuucaguuguaaauauucagaaagagauagaucgccugaaugagguggcuaagaaccugaacgaaucucucauugaucuccaagagcugggaaaguacgaacaauacaucaaauggccuucugggcgucgccgaagacgacgaggguccggcggcucagggagcggcuauaucccugaggcgccucgggacggacaagcuuaugugaggaaagauggagaauggguauugcugucaaccuuccugggauaauga (SEQ ID NO: 93)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSGIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIAGTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHAGQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSGPNDILSRLPKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTDVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 94) WuS_IgE_StrepHis_pVaxgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcaagccaatgcgtgaatctgactacaaggactcagctgccccctgcatacacgaacagtttcacccgcggtgtatattatccggacaaagtattcaggtctagtgtgctgcactcaacccaggatttgtttctgcccttcttctctaacgtgacatggttccacgccatccatgtgtcaggtacgaacggtaccaagagatttgataacccggtactgccatttaatgacggcgtctattttgcttccactgagaagagcaacatcataagaggctggatctttggaactactctggacagcaaaacccagagcttgctgatcgtgaacaacgcgacaaatgtagtgatcaaagtatgtgagtttcaattctgtaacgatcccttccttggggtttattaccataagaataataagagttggatggagtccgaatttagagtttactcctcagctaataactgtacgttcgagtatgtctcccaaccttttcttatggatctcgaagggaaacagggtaactttaagaatcttcgagaatttgtgttcaagaacatcgacggttattttaagatctacagtaagcatactcccataaatctggttagagatctcccgcaaggattttccgcactggagccccttgtagaccttcccattggaataaacataacacgtttccagacactcctcgctctgcataggtcatatctcaccccgggcgattcttccagcggatggaccgctggagctgctgcttactacgtaggatacctgcaaccccggacatttctgctcaagtataacgaaaatgggactattacggacgctgtggactgtgctcttgacccacttagcgagacaaaatgcacgctgaaaagttttaccgtggagaaggggatctatcaaacgagcaattttagggttcagcctaccgaatcaatcgtcagatttcccaatatcactaacctgtgccctttcggggaagttttcaacgcaacccggtttgcgagcgtatacgcttggaatcgcaaaaggataagcaattgcgttgccgattactccgttctttacaattcagcatcattttctacttttaaatgctacggcgtgtctcccacaaaactgaatgacctgtgttttacgaacgtgtatgcagacagctttgtgattaggggtgatgaggttagacaaatcgcaccaggtcagaccggtaagatcgctgattacaactacaaactgcccgatgacttcacaggatgcgtgattgcctggaattccaacaatctggattctaaggttggcggcaattacaattacctgtataggttgtttcggaagtcaaacctgaaacccttcgaaagagacatttctaccgagatttatcaagcgggttcaactccttgtaatggagttgaaggcttcaattgttactttccccttcaatcatacggattccaaccaaccaatggggtcggataccagccatatagggttgttgtcctgtcattcgaacttctccacgcaccagccaccgtatgtggacccaagaagtctactaatctggtgaagaacaaatgcgtcaatttcaactttaatgggttgaccggcactggggtgctgactgaatccaacaagaagtttctgccgttccaacaattcggacgcgatatcgctgatacaaccgatgccgttagagatccccaaacattggagattctggatattacgccttgttcattcggtggtgtttccgtgattacccctggcaccaatacgagtaaccaagtggcggtgctgtatcaagatgtgaactgtactgaagtgccggtggctatacatgccgaccaactcacaccaacatggagagtatatagcacgggttccaatgtgtttcaaactagggctggctgtttgattggcgctgaacatgttaataattcctatgaatgcgatattcccatcggtgccgggatttgcgcaagttatcaaacgcaaactaactcccccgggtcagcatcctctgtcgcttcccaatcaatcatcgcctataccatgagtcttggggcagaaaattccgttgcttattctaacaattccattgcaattcctacgaacttcaccatctcagttactacagaaatacttcccgtgtcaatgacgaagacatccgtagattgcacaatgtatatatgtggggactcaactgaatgctcaaacctgctcctgcaatacggatcattttgcacccaactgaacagagcattgaccggtatagccgtggagcaagataagaacactcaagaagtattcgcccaggtcaaacaaatctataaaactccgcctataaaagattttggcggctttaacttttcccaaatactgcctgacccaagtaagccctcaaaacgtagctttatagaggacctcttgtttaataaggtgacactcgctgacgctggattcattaagcaatatggtgactgcttgggagatattgccgcccgcgatctcatttgtgcacaaaagttcaacggcctcacagtcctgccccctctgctgacggatgaaatgatcgctcaatacacctcagctctcctggcaggcaccataacaagcgggtggacatttggtgccggggcagcactgcaaatcccattcgcaatgcaaatggcttataggttcaatgggatcggcgtaactcaaaatgtcctctacgagaaccagaaactcatagctaaccaattcaattctgcaatcgggaaaatccaggactccctgagctcaacggccagcgcactgggcaagctccaagatgtggtcaaccaaaacgcacaagcactgaatactcttgtgaaacaactcagctccaatttcggggcaatatcaagtgtcctcaatgatattcttagcaggcttgatccacccgaagccgaggtgcagatcgacaggctcataacaggcaggctccagtcccttcaaacgtatgtaactcagcaactgattcgggctgccgagattcgagcttcagctaatttggcagctacgaagatgagcgaatgcgtcctgggacagtctaaaagagtagacttttgcggcaaagggtatcatctgatgagcttcccacaaagtgctccacatggcgtggttttcctgcatgtcacttatgttcccgcacaagagaagaacttcactaccgcaccagcgatctgtcacgatggtaaagcacatttcccgcgggaaggcgtattcgtatctaacggcacccactggttcgttactcaacgcaacttttatgaaccacaaatcattacaaccgataacacttttgtttcaggcaattgcgatgttgtcatcggcattgtgaataacactgtgtacgatccacttcaaccagaattggacagctttaaagaggagcttgataagtatttcaagaatcatacctctcccgacgtggacctcggggacatctctggaataaatgctagcgtcgttaatatacagaaagagattgatcgtctgaacgaagtggctaagaatctgaatgaaagccttatcgatctgcaagaactggggaagtacgaacagggatacataccggaagccccacgcgacggtcaggcttatgttaggaaggatggagaatgggttttgctctccacgtttctcgggcttgaagttttgttccaaggaccctggtcacacccccaatttgagaaacaccatcaccaccatcaccaccactgataa (SEQ ID NO: 95)gccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaagccaaugcgugaaucugacuacaaggacucagcugcccccugcauacacgaacaguuucacccgcgguguauauuauccggacaaaguauucaggucuagugugcugcacucaacccaggauuuguuucugcccuucuucucuaacgugacaugguuccacgccauccaugugucagguacgaacgguaccaagagauuugauaacccgguacugccauuuaaugacggcgucuauuuugcuuccacugagaagagcaacaucauaagaggcuggaucuuuggaacuacucuggacagcaaaacccagagcuugcugaucgugaacaacgcgacaaauguagugaucaaaguaugugaguuucaauucuguaacgaucccuuccuugggguuuauuaccauaagaauaauaagaguuggauggaguccgaauuuagaguuuacuccucagcuaauaacuguacguucgaguaugucucccaaccuuuucuuauggaucucgaagggaaacaggguaacuuuaagaaucuucgagaauuuguguucaagaacaucgacgguuauuuuaagaucuacaguaagcauacucccauaaaucugguuagagaucucccgcaaggauuuuccgcacuggagccccuuguagaccuucccauuggaauaaacauaacacguuuccagacacuccucgcucugcauaggucauaucucaccccgggcgauucuuccagcggauggaccgcuggagcugcugcuuacuacguaggauaccugcaaccccggacauuucugcucaaguauaacgaaaaugggacuauuacggacgcuguggacugugcucuugacccacuuagcgagacaaaaugcacgcugaaaaguuuuaccguggagaaggggaucuaucaaacgagcaauuuuaggguucagccuaccgaaucaaucgucagauuucccaauaucacuaaccugugcccuuucggggaaguuuucaacgcaacccgguuugcgagcguauacgcuuggaaucgcaaaaggauaagcaauugcguugccgauuacuccguucuuuacaauucagcaucauuuucuacuuuuaaaugcuacggcgugucucccacaaaacugaaugaccuguguuuuacgaacguguaugcagacagcuuugugauuaggggugaugagguuagacaaaucgcaccaggucagaccgguaagaucgcugauuacaacuacaaacugcccgaugacuucacaggaugcgugauugccuggaauuccaacaaucuggauucuaagguuggcggcaauuacaauuaccuguauagguuguuucggaagucaaaccugaaacccuucgaaagagacauuucuaccgagauuuaucaagcggguucaacuccuuguaauggaguugaaggcuucaauuguuacuuuccccuucaaucauacggauuccaaccaaccaauggggucggauaccagccauauaggguuguuguccugucauucgaacuucuccacgcaccagccaccguauguggacccaagaagucuacuaaucuggugaagaacaaaugcgucaauuucaacuuuaauggguugaccggcacuggggugcugacugaauccaacaagaaguuucugccguuccaacaauucggacgcgauaucgcugauacaaccgaugccguuagagauccccaaacauuggagauucuggauauuacgccuuguucauucggugguguuuccgugauuaccccuggcaccaauacgaguaaccaaguggcggugcuguaucaagaugugaacuguacugaagugccgguggcuauacaugccgaccaacucacaccaacauggagaguauauagcacggguuccaauguguuucaaacuagggcuggcuguuugauuggcgcugaacauguuaauaauuccuaugaaugcgauauucccaucggugccgggauuugcgcaaguuaucaaacgcaaacuaacucccccgggucagcauccucugucgcuucccaaucaaucaucgccuauaccaugagucuuggggcagaaaauuccguugcuuauucuaacaauuccauugcaauuccuacgaacuucaccaucucaguuacuacagaaauacuucccgugucaaugacgaagacauccguagauugcacaauguauauauguggggacucaacugaaugcucaaaccugcuccugcaauacggaucauuuugcacccaacugaacagagcauugaccgguauagccguggagcaagauaagaacacucaagaaguauucgcccaggucaaacaaaucuauaaaacuccgccuauaaaagauuuuggcggcuuuaacuuuucccaaauacugccugacccaaguaagcccucaaaacguagcuuuauagaggaccucuuguuuaauaaggugacacucgcugacgcuggauucauuaagcaauauggugacugcuugggagauauugccgcccgcgaucucauuugugcacaaaaguucaacggccucacaguccugcccccucugcugacggaugaaaugaucgcucaauacaccucagcucuccuggcaggcaccauaacaagcggguggacauuuggugccggggcagcacugcaaaucccauucgcaaugcaaauggcuuauagguucaaugggaucggcguaacucaaaauguccucuacgagaaccagaaacucauagcuaaccaauucaauucugcaaucgggaaaauccaggacucccugagcucaacggccagcgcacugggcaagcuccaagauguggucaaccaaaacgcacaagcacugaauacucuugugaaacaacucagcuccaauuucggggcaauaucaaguguccucaaugauauucuuagcaggcuugauccacccgaagccgaggugcagaucgacaggcucauaacaggcaggcuccagucccuucaaacguauguaacucagcaacugauucgggcugccgagauucgagcuucagcuaauuuggcagcuacgaagaugagcgaaugcguccugggacagucuaaaagaguagacuuuugcggcaaaggguaucaucugaugagcuucccacaaagugcuccacauggcgugguuuuccugcaugucacuuauguucccgcacaagagaagaacuucacuaccgcaccagcgaucugucacgaugguaaagcacauuucccgcgggaaggcguauucguaucuaacggcacccacugguucguuacucaacgcaacuuuuaugaaccacaaaucauuacaaccgauaacacuuuuguuucaggcaauugcgauguugucaucggcauugugaauaacacuguguacgauccacuucaaccagaauuggacagcuuuaaagaggagcuugauaaguauuucaagaaucauaccucucccgacguggaccucggggacaucucuggaauaaaugcuagcgucguuaauauacagaaagagauugaucgucugaacgaaguggcuaagaaucugaaugaaagccuuaucgaucugcaagaacuggggaaguacgaacagggauacauaccggaagccccacgcgacggucaggcuuauguuaggaaggauggagaauggguuuugcucuccacguuucucgggcuugaaguuuuguuccaaggacccuggucacacccccaauuugagaaacaccaucaccaccaucaccaccacugauaa(SEQ ID NO: 96) MDWTWILFLVAAATRVHSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPGSASSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISS VLNDILSRLD

EAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQGYIPEAPRDGQAYVRKDGEWVLLSTFLGLEVLFQGPWSHPQFEKHHHHHHHH** (SEQ ID NO: 97)WuS_IgE_DownDS2_2P_pVaxgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcatcacagtgcgttaatctgaccacccgtacacaactcccacccgcatacacaaatagctttacacgcggagtgtattaccccgataaagtctttcggagctcagtgctccattctactcaagatcttttcctgccgttctttagtaacgttacttggtttcatgcaatacatgtgtctggcacaaacggaaccaaacgttttgataatccggtgttgccatttaatgatggtgtatattttgcttccacggaaaagtcaaacatcatccgtgggtggatctttggcaccactcttgatagcaaatgtcaaagccttctgattgttaataacgctacaaacgtcgtaattaaagtgtgtgaattccagttctgtaatgaccccttcctcggagtatattaccacaagaataacaaatcttggatggagagcgaatttagagtttacagttcagccaataactgtacatttgaatatgtcagtcagcctttcctcatggacctcgaaggtaaacaaggtaattttaagaacttgagagagttcgtgtttaagaacatcgatggctatttcaaaatttactctaagcacacaccaatcaacctggttcgagacctgccccagggtttctcagctttggaaccattggtggacctgccaatcggcattaacattaccagatttcaaactttgttggcactccaccggtcatatcttacccccggagacagttcctcaggctggacggcaggcgccgccgcgtactatgttgggtatctccaaccccgaaccttccttctcaaatacaatgaaaacgggacgattacagatgcagtcgattgcgccctggaccccttgtccgaaactaaatgcactctgaagagtttcacggtagagaagggaatctatcaaacgagcaattttcgagtccaaccaacggaatctattgtgcggtttcccaatatcacaaacctctgtccattcggagaagtctttaatgctaccaggtttgcgtctgtatatgcatggaaccgaaagaggatttccaattgcgtagcggactacagtgtcctttataacagcgcttcattttccacgtttaagtgttatggtgtttctccaacgaaactcaacgacctctgttttactaacgtttacgctgacagctttgttatacgtggggacgaagtcaggcaaattgctcctggacagactggaaagatcgctgattataattataaacttcctgacgatttcaccggctgcgttattgcatggaactccaacaatctggattcaaaagtgggtggaaattataattatctgtataggttgtttcggaagagcaatcttaagccctttgagcgggacatatgtaccgaaatttaccaagcaggctccaccccatgcaatggagtagaagggttcaattgctattttcctctgcaaagttatggctttcaacccaccaacggagttgggtatcaaccttacagggttgtcgtgctgagtttcgaattgctccacgcacccgctacagtatgtggccccaagaagtccactaatcttgttaagaataaatgcgtgaacttcaacttcaatggacttacaggtactggagtactcacggaatcaaacaagaaatttctcccatttcaacagtttggccgagatatagctgacaccacagatgctgttcgcgacccccagacgttggaaatacttgatatcactccctgcagcttcggcggcgtgagcgtgatcactccaggtactaatacgagcaatcaagttgccgttctgtaccaagatgtgaactgcaccgaggttccagtggcaattcacgccgaccaacttactcccacctggcgggtctattccaccggatcaaacgtcttccaaactcgcgctggttgccttatcggtgcagagcacgttaataattcctatgaatgtgacattcccataggagcaggcatctgtgcatcttatcaaacccagactaattcccctggttccgcttcctctgttgcatcccagtccataattgcctacactatgagtctcggggctgaaaattccgtggcctattctaataattcaatcgccatcccaaccaattttaccatatccgtaacgactgaaatacttcctgtcagtatgaccaagacctcagtggactgcaccatgtacatctgcggcgattctactgaatgttccaatctgcttttgcaatatggttcattctgcacccaactcaacagggctcttacagggatcgccgtcgaacaggataagaatacccaggaagtgttcgcccaagttaagcaaatttacaagacaccacccatcaaggacttcggcgggttcaacttcagccaaattctgcccgacccgtctaagccttctaagcgctctttcattgaggatcttttgttcaataaggttacgcttgccgatgcagggtttatcaaacagtatggcgactgtcttggggatatcgcagctagggatcttatttgtgcacagaaatttaatggcctgactgttcttccccctttgctcactgacgagatgattgcccagtacacttcagctctcctggccgggactataacttctggttggaccttcggagctggcgccgccctgcaaattccatttgcaatgcagatggcttatcgcttcaacggaattggggtgacccaaaatgttctctacgagaaccagaaactcattgcaaaccagttcaattctgcgatcgggaagatccaggattccctgtctagtacggctagtgccctcggtaagctccaagacgtcgtcaaccaaaacgcccaggccttgaacacccttgtcaaacaactgagctccaattttggggctattagcagtgtgctgaatgatatcctgtcccgccttgacccaccggaagcggaagtccaaattgatcgactgatcactgggcgtctccaatcccttcaaacttacgtgacccaacaactcatccgagcagctgagattagggctagcgctaaccttgctgctactaagatgtcagagtgtgtcctcggccagtctaagagagtggacttttgtgggaaagggtaccacttgatgtcattcccacaaagcgccccacacggcgtggtgtttctccacgtcacttacgttccagctcaggaaaagaactttaccaccgcccccgctatatgtcatgatgggaaggcccactttcctcgtgaaggtgtctttgtcagcaatggcacacactggtttgtgacccaacggaatttctatgagcctcagattattaccacggataacactttcgtatcagggaattgtgatgtggttatcggcatcgttaataatacagtgtatgacccactgcagccagagcttgacagcttcaaagaagagctcgataagtactttaagaatcatacaagtcctgacgttgatcttggggatattagtgggattaacgccagcgtcgtcaatattcagaaagagattgacaggttgaacgaagtagctaagaatcttaatgaaagcctgatagatttgcaagaacttggtaagtatgagcaggggtacatacccgaggctcctcgggatgggcaggcctatgtacgcaaagacggtgaatgggtattgctcagcacttttctcggctgataa (SEQ ID NO: 98)gccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaucacagugcguuaaucugaccacccguacacaacucccacccgcauacacaaauagcuuuacacgcggaguguauuaccccgauaaagucuuucggagcucagugcuccauucuacucaagaucuuuuccugccguucuuuaguaacguuacuugguuucaugcaauacaugugucuggcacaaacggaaccaaacguuuugauaauccgguguugccauuuaaugaugguguauauuuugcuuccacggaaaagucaaacaucauccguggguggaucuuuggcaccacucuugauagcaaaugucaaagccuucugauuguuaauaacgcuacaaacgucguaauuaaagugugugaauuccaguucuguaaugaccccuuccucggaguauauuaccacaagaauaacaaaucuuggauggagagcgaauuuagaguuuacaguucagccaauaacuguacauuugaauaugucagucagccuuuccucauggaccucgaagguaaacaagguaauuuuaagaacuugagagaguucguguuuaagaacaucgauggcuauuucaaaauuuacucuaagcacacaccaaucaaccugguucgagaccugccccaggguuucucagcuuuggaaccauugguggaccugccaaucggcauuaacauuaccagauuucaaacuuuguuggcacuccaccggucauaucuuacccccggagacaguuccucaggcuggacggcaggcgccgccgcguacuauguuggguaucuccaaccccgaaccuuccuucucaaauacaaugaaaacgggacgauuacagaugcagucgauugcgcccuggaccccuuguccgaaacuaaaugcacucugaagaguuucacgguagagaagggaaucuaucaaacgagcaauuuucgaguccaaccaacggaaucuauugugcgguuucccaauaucacaaaccucuguccauucggagaagucuuuaaugcuaccagguuugcgucuguauaugcauggaaccgaaagaggauuuccaauugcguagcggacuacaguguccuuuauaacagcgcuucauuuuccacguuuaaguguuaugguguuucuccaacgaaacucaacgaccucuguuuuacuaacguuuacgcugacagcuuuguuauacguggggacgaagucaggcaaauugcuccuggacagacuggaaagaucgcugauuauaauuauaaacuuccugacgauuucaccggcugcguuauugcauggaacuccaacaaucuggauucaaaaguggguggaaauuauaauuaucuguauagguuguuucggaagagcaaucuuaagcccuuugagcgggacauauguaccgaaauuuaccaagcaggcuccaccccaugcaauggaguagaaggguucaauugcuauuuuccucugcaaaguuauggcuuucaacccaccaacggaguuggguaucaaccuuacaggguugucgugcugaguuucgaauugcuccacgcacccgcuacaguauguggccccaagaaguccacuaaucuuguuaagaauaaaugcgugaacuucaacuucaauggacuuacagguacuggaguacucacggaaucaaacaagaaauuucucccauuucaacaguuuggccgagauauagcugacaccacagaugcuguucgcgacccccagacguuggaaauacuugauaucacucccugcagcuucggcggcgugagcgugaucacuccagguacuaauacgagcaaucaaguugccguucuguaccaagaugugaacugcaccgagguuccaguggcaauucacgccgaccaacuuacucccaccuggcgggucuauuccaccggaucaaacgucuuccaaacucgcgcugguugccuuaucggugcagagcacguuaauaauuccuaugaaugugacauucccauaggagcaggcaucugugcaucuuaucaaacccagacuaauuccccugguuccgcuuccucuguugcaucccaguccauaauugccuacacuaugagucucggggcugaaaauuccguggccuauucuaauaauucaaucgccaucccaaccaauuuuaccauauccguaacgacugaaauacuuccugucaguaugaccaagaccucaguggacugcaccauguacaucugcggcgauucuacugaauguuccaaucugcuuuugcaauaugguucauucugcacccaacucaacagggcucuuacagggaucgccgucgaacaggauaagaauacccaggaaguguucgcccaaguuaagcaaauuuacaagacaccacccaucaaggacuucggcggguucaacuucagccaaauucugcccgacccgucuaagccuucuaagcgcucuuucauugaggaucuuuuguucaauaagguuacgcuugccgaugcaggguuuaucaaacaguauggcgacugucuuggggauaucgcagcuagggaucuuauuugugcacagaaauuuaauggccugacuguucuucccccuuugcucacugacgagaugauugcccaguacacuucagcucuccuggccgggacuauaacuucugguuggaccuucggagcuggcgccgcccugcaaauuccauuugcaaugcagauggcuuaucgcuucaacggaauuggggugacccaaaauguucucuacgagaaccagaaacucauugcaaaccaguucaauucugcgaucgggaagauccaggauucccugucuaguacggcuagugcccucgguaagcuccaagacgucgucaaccaaaacgcccaggccuugaacacccuugucaaacaacugagcuccaauuuuggggcuauuagcagugugcugaaugauauccugucccgccuugacccaccggaagcggaaguccaaauugaucgacugaucacugggcgucuccaaucccuucaaacuuacgugacccaacaacucauccgagcagcugagauuagggcuagcgcuaaccuugcugcuacuaagaugucagaguguguccucggccagucuaagagaguggacuuuugugggaaaggguaccacuugaugucauucccacaaagcgccccacacggcgugguguuucuccacgucacuuacguuccagcucaggaaaagaacuuuaccaccgcccccgcuauaugucaugaugggaaggcccacuuuccucgugaaggugucuuugucagcaauggcacacacugguuugugacccaacggaauuucuaugagccucagauuauuaccacggauaacacuuucguaucagggaauugugaugugguuaucggcaucguuaauaauacaguguaugacccacugcagccagagcuugacagcuucaaagaagagcucgauaaguacuuuaagaaucauacaaguccugacguugaucuuggggauauuagugggauuaacgccagcgucgucaauauucagaaagagauugacagguugaacgaaguagcuaagaaucuuaaugaaagccugauagauuugcaagaacuugguaaguaugagcagggguacauacccgaggcuccucgggaugggcaggccuauguacgcaaagacggugaauggguauugcucagcacuuuucucggcugauaa (SEQ ID NO: 99)MDWTWILFLVAAATRVHSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGT TLDSK

QSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDI

TEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPGSASSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISS VLNDILSRLD

EAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 100)WuS_IgE_DownDS1 _2P_pVaxgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcatcccagtgcgtgaacctgaccacccgaactcaactcccaccagcatacaccaactcatttacaagaggagtttattacccggacaaggtatttcgaagttcagttcttcacagcacccaagacctgtttctgccattcttcagtaatgtcacttggtttcacgcgatacatgtcagcggtacaaacgggacaaagcgattcgataacccagtactcccattcaacgacggagtgtattttgcatctacagagaaatccaacattatacgcgggtggatctttggaactactctggactccaagacacagagcctgctcattgtgaacaatgcaacgaatgtcgtcataaaagtctgtgaatttcaattttgcaacgatcctttcctcggagtctattaccataagaacaataagagttggatggagagtgagtttcgcgtctattcttccgcgaacaattgtacatttgaatatgtatcacaaccctttcttatggatttggaaggcaaacaaggtaacttcaagaacttgcgcgagttcgtgttcaagaacatagactgttattttaagatctatagtaagcatacgccaatcaatctggtgcgagatttgcctcagggcttttctgctcttgaacccttggttgatctgcccatcgggatcaacataaccagatttcaaacgttgctcgcactccaccgcagctatctcactcctggcgattcctcatctgggtggaccgccggagctgctgcttattacgtcggctatctccagccgcgtactttcctgctcaagtataatgagaatggcaccattaccgatgctgtggattgtgctcttgatccactctctgaaaccaaatgcactctcaagtcttttaccgtggaaaagggtatttatcagacatctaattttcgggtgcaacctactgagtcaattgtacggtttcctaacataactaacctttgtccatttggggaagtcttcaatgccacgcggttcgcatcagtctatgcatggaacagaaaacgtatctccaactgcgtcgccgattattccgtcctttacaatagcgctagcttttccacattcaaatgttatggcgtatcaccaaccaaacttaacgatctctgctttactaatgtctacgctgactctttcgttattcgaggtgacgaggtgcgccaaattgcgcctggtcaaaccggaaagattgccgattataactacaagctccccgacgactttacgggttgtgtgatcgcctggaatagcaataacctcgattctaaagttggcggtaattataactatctgtacagactctttaggaaaagtaatctcaagcccttttgcagggatatctcaaccgaaatctaccaagccggcagcactccttgcaatggtgtcgaggggtttaattgttatttcccactgcaatcttacggctttcaaccgactaatggagtcggttatcaaccctatagggtggtggtactctcctttgaacttttgcacgctccggcaacagtttgtggaccaaagaaaagtacgaaccttgttaagaataagtgtgttaatttcaattttaacggcctcactggaacaggtgtcctcacagaaagcaacaagaagtttctccctttccaacagtttggacgggatatcgccgacactactgacgccgtcagagatcctcaaactctcgaaatcttggatatcacaccatgttctttcggtggtgtctccgtcataacaccaggaactaacacctctaatcaagtggccgtgctctatcaggacgtcaattgcacagaagtgcctgtcgcaatccatgctgatcagctcactcccacctggcgtgtgtattccactggctctaatgtctttcagacacgggcaggttgccttattggggcagagcatgtgaacaattcctacgaatgcgatatacccattggggcaggcatttgcgccagctaccaaacccaaactaacagccccgggagtgccagcagcgtggcatctcagtccattattgcctatacgatgagcctgggtgctgaaaatagcgtggcttatagtaataactctatcgccatacccacaaacttcaccatttcagtgaccaccgaaatccttcctgtttctatgaccaaaacgtccgtcgattgtacaatgtacatttgcggcgatagcactgaatgttcaaacctgctcctgcaatacggctctttctgtactcagctcaaccgggcactcaccggcatagccgtcgaacaagacaagaatacccaggaagtctttgcgcaggtgaaacaaatctataagaccccaccaataaaagatttcggcggttttaatttcagccaaatcttgcctgatcccagcaagccatctaaacggtctttcattgaagatctcctgttcaacaaggttacgctggctgacgccgggtttattaagcaatatggcgattgccttggggacattgccgcacgagacctcatttgtgcccagaaattcaacgggctcaccgtattgcccccgctcctcacagacgaaatgatcgcccaatatacaagcgccctgcttgcgggaaccattacaagcggttggacctttggtgccggcgcagctctgcaaatacccttcgcaatgcaaatggcatatcggtttaatggaattggcgtaacccaaaacgtgctgtatgaaaaccagaaactgatcgcaaatcaattcaatagtgctataggaaagatccaagacagtctgtcttccactgctagcgcgctggggaagctccaagacgttgtgaaccaaaacgcgcaggccctgaataccctggtgaagcaactttcaagcaatttcggtgctatatcttctgtcctcaatgacattctctctcggctcgatcccccggaagccgaagttcagatagaccgtttgatcacaggccgcttgcaatccctgcaaacctacgttacacaacaactgattcgcgccgccgaaattcgggcatccgccaatctggccgcaaccaaaatgtccgagtgtgttctcggtcaatccaaacgcgtggatttctgcggaaaaggataccatttgatgtcatttccacaatcagctccacacggtgttgtattcctgcacgtgacctacgtgccagcccaggagaagaattttactactgcgcccgccatttgtcatgacgggaaggctcattttcctcgggaaggggttttcgtctcaaacggtacccattggttcgtgactcagaggaacttttatgaacctcaaatcataacgaccgataacacgtttgtaagtggcaattgcgacgtggtcatcgggattgtaaacaatactgtctatgaccctctccaaccagagcttgacagctttaaagaagagcttgataaatactttaagaaccatacctcaccagacgtcgatttgggagatatcagtggcattaatgcctctgtcgtcaatatccagaaagagattgaccgcttgaacgaagttgccaagaatcttaatgagtctctgattgacttgcaagaattgggaaaatatgaacaaggatatattccagaagcccctcgcgatgggcaagcatatgttcgaaaggatggggaatgggtgctgctcagcacctttctcggttgataa (SEQ ID NO: 101)gccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaucccagugcgugaaccugaccacccgaacucaacucccaccagcauacaccaacucauuuacaagaggaguuuauuacccggacaagguauuucgaaguucaguucuucacagcacccaagaccuguuucugccauucuucaguaaugucacuugguuucacgcgauacaugucagcgguacaaacgggacaaagcgauucgauaacccaguacucccauucaacgacggaguguauuuugcaucuacagagaaauccaacauuauacgcggguggaucuuuggaacuacucuggacuccaagacacagagccugcucauugugaacaaugcaacgaaugucgucauaaaagucugugaauuucaauuuugcaacgauccuuuccucggagucuauuaccauaagaacaauaagaguuggauggagagugaguuucgcgucuauucuuccgcgaacaauuguacauuugaauauguaucacaacccuuucuuauggauuuggaaggcaaacaagguaacuucaagaacuugcgcgaguucguguucaagaacauagacuguuauuuuaagaucuauaguaagcauacgccaaucaaucuggugcgagauuugccucagggcuuuucugcucuugaacccuugguugaucugcccaucgggaucaacauaaccagauuucaaacguugcucgcacuccaccgcagcuaucucacuccuggcgauuccucaucuggguggaccgccggagcugcugcuuauuacgucggcuaucuccagccgcguacuuuccugcucaaguauaaugagaauggcaccauuaccgaugcuguggauugugcucuugauccacucucugaaaccaaaugcacucucaagucuuuuaccguggaaaaggguauuuaucagacaucuaauuuucgggugcaaccuacugagucaauuguacgguuuccuaacauaacuaaccuuuguccauuuggggaagucuucaaugccacgcgguucgcaucagucuaugcauggaacagaaaacguaucuccaacugcgucgccgauuauuccguccuuuacaauagcgcuagcuuuuccacauucaaauguuauggcguaucaccaaccaaacuuaacgaucucugcuuuacuaaugucuacgcugacucuuucguuauucgaggugacgaggugcgccaaauugcgccuggucaaaccggaaagauugccgauuauaacuacaagcuccccgacgacuuuacggguugugugaucgccuggaauagcaauaaccucgauucuaaaguuggcgguaauuauaacuaucuguacagacucuuuaggaaaaguaaucucaagcccuuuugcagggauaucucaaccgaaaucuaccaagccggcagcacuccuugcaauggugucgagggguuuaauuguuauuucccacugcaaucuuacggcuuucaaccgacuaauggagucgguuaucaacccuauaggguggugguacucuccuuugaacuuuugcacgcuccggcaacaguuuguggaccaaagaaaaguacgaaccuuguuaagaauaaguguguuaauuucaauuuuaacggccucacuggaacagguguccucacagaaagcaacaagaaguuucucccuuuccaacaguuuggacgggauaucgccgacacuacugacgccgucagagauccucaaacucucgaaaucuuggauaucacaccauguucuuucgguggugucuccgucauaacaccaggaacuaacaccucuaaucaaguggccgugcucuaucaggacgucaauugcacagaagugccugucgcaauccaugcugaucagcucacucccaccuggcguguguauuccacuggcucuaaugucuuucagacacgggcagguugccuuauuggggcagagcaugugaacaauuccuacgaaugcgauauacccauuggggcaggcauuugcgccagcuaccaaacccaaacuaacagccccgggagugccagcagcguggcaucucaguccauuauugccuauacgaugagccugggugcugaaaauagcguggcuuauaguaauaacucuaucgccauacccacaaacuucaccauuucagugaccaccgaaauccuuccuguuucuaugaccaaaacguccgucgauuguacaauguacauuugcggcgauagcacugaauguucaaaccugcuccugcaauacggcucuuucuguacucagcucaaccgggcacucaccggcauagccgucgaacaagacaagaauacccaggaagucuuugcgcaggugaaacaaaucuauaagaccccaccaauaaaagauuucggcgguuuuaauuucagccaaaucuugccugaucccagcaagccaucuaaacggucuuucauugaagaucuccuguucaacaagguuacgcuggcugacgccggguuuauuaagcaauauggcgauugccuuggggacauugccgcacgagaccucauuugugcccagaaauucaacgggcucaccguauugcccccgcuccucacagacgaaaugaucgcccaauauacaagcgcccugcuugcgggaaccauuacaagcgguuggaccuuuggugccggcgcagcucugcaaauacccuucgcaaugcaaauggcauaucgguuuaauggaauuggcguaacccaaaacgugcuguaugaaaaccagaaacugaucgcaaaucaauucaauagugcuauaggaaagauccaagacagucugucuuccacugcuagcgcgcuggggaagcuccaagacguugugaaccaaaacgcgcaggcccugaauacccuggugaagcaacuuucaagcaauuucggugcuauaucuucuguccucaaugacauucucucucggcucgaucccccggaagccgaaguucagauagaccguuugaucacaggccgcuugcaaucccugcaaaccuacguuacacaacaacugauucgcgccgccgaaauucgggcauccgccaaucuggccgcaaccaaaauguccgaguguguucucggucaauccaaacgcguggauuucugcggaaaaggauaccauuugaugucauuuccacaaucagcuccacacgguguuguauuccugcacgugaccuacgugccagcccaggagaagaauuuuacuacugcgcccgccauuugucaugacgggaaggcucauuuuccucgggaagggguuuucgucucaaacgguacccauugguucgugacucagaggaacuuuuaugaaccucaaaucauaacgaccgauaacacguuuguaaguggcaauugcgacguggucaucgggauuguaaacaauacugucuaugacccucuccaaccagagcuugacagcuuuaaagaagagcuugauaaauacuuuaagaaccauaccucaccagacgucgauuugggagauaucaguggcauuaaugccucugucgucaauauccagaaagagauugaccgcuugaacgaaguugccaagaaucuuaaugagucucugauugacuugcaagaauugggaaaauaugaacaaggauauauuccagaagccccucgcgaugggcaagcauauguucgaaaggauggggaaugggugcugcucagcaccuuucucgguugauaa (SEQ ID NO: 102)MDWTWILFLVAAATRVHSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNID

YFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFCRDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPGSASSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 103) WuS_IgE_2P_UpGly_pVaxgccaccatggattggacctggatacttttcctcgtggccgcagcaacaagagtccactcctctcagtgcgttaacctgactactagaacccaattgcccccggcatacacaaactctttcacccggggtgtctactatcccgacaaagtgtttagaagtagcgtgctgcacagcacccaagatctctttctgccattcttctcaaacgtcacctggtttcacgccatccatgtaagcgggaccaacggcacaaagcgttttgataaccctgttttgccattcaatgatggcgtgtattttgcttccactgagaaaagcaacatcattagagggtggatatttggcacaacgcttgactccaagacgcagagtcttttgatagtaaacaacgcaactaatgtggtcattaaagtctgtgaatttcaattttgcaatgaccccttccttggagtctattaccacaagaacaacaaaagctggatggaaagcgaatttagggtctacagctctgccaataactgcacattcgaatacgtcagccaaccattcttgatggacctggaaggcaagcaaggaaactttaagaatctgagggaatttgtgtttaagaatatcgacggatattttaagatctattccaagcatactcccattaatctcgttcgtgaccttcctcagggtttctctgcattggaacccctcgtagatttgcccattgggattaatatcactagattccagacgctgcttgcactccatcgatcttatctgacccctggtgactcctcttccgggtggacggcgggtgctgcagcctactacgttggctatttgcaacctaggacctttctgttgaagtataatgagaatgggactattactgatgccgttgattgcgccctcgatccgctgtcagaaacaaagtgcaccctgaagagcttcacagtagaaaagggaatctatcaaacctcaaatttccgcgttcaaccaactgaatcaatcgtgcgttttcctaacatcacaaatctgtgtccgtttggagaagtatttaatgcgacgcgtttcgcaagcgtctacgcgtggaatcgcaaacgtatctctaattgcgtagcagattattctgtgctgtacaatagcgcatctttctcaacgtttaagtgctacggcgttaatgggaccaagctgaatgatctctgtttcactaatgtgtacgcagacagttttgtaattagaggagacgaggttaggcaaatagcaccgggtcaaactggcaaaatcgccgactataactacaagctccctgatgacttcacgggctgcgtaattgcttggaactctaataacctggactctaaagtcggcgggaattataattatctctatcggttgtttcgaaaatccaatctcaaaccctttgagcgggacatcaatactacaatttatcaagctggtagtactccttgcaatggggtagaaggcttcaattgttatttcccccttcaatcttacggatttcaacccacgaacggcgtagggtaccagccctatcgagtggtggtactgtcattcgaacttaatcacgccccagcaacagtctgcgggcctaagaaaagcacgaatcttgtcaagaataagtgtgtaaatttcaacttcaatggtcttacaggcacgggagtgctcactgagtctaataagaaatttcttcctttccaacaattcggtcgtgatattgccgatactactgatgcagtccgagatccacaaactctcgaaatcctcgatattactccttgtagttttggcggcgtctccgtgatcaccccagggaccaacactagtaaccaagtggcggtgctctaccaagatgttaactgcacagaagtcccggtagcgatccatgccgaccagctcactcccacatggcgtgtttacagcacagggtcaaacgttttccagacccgtgccggatgtcttataggagccgaacacgtaaataacagttatgaatgcgatatcccaattggtgcaggtatctgtgcgtcatatcaaacccaaactaattctccggggtccgcctcaagcgttgcctcacaatcaataatcgcctacacaatgtccctcggtgccgaaaattcagtcgcttactctaacaatagcattgctatccctaccaacttcactatttctgttaccacggaaattttgcctgtatccatgaccaaaacatctgttgattgcacgatgtacatctgcggggattctaccgaatgttctaacctgcttctgcaatacggctccttctgcacccaattgaaccgcgcactgactgggattgctgtggaacaagacaagaatactcaagaagtatttgcccaggtcaaacagatttacaaaactcccccaattaaagatttcggcggtttcaattttagtcaaattctgccagatccaagtaagccatccaaacgctcatttattgaggacctgctctttaataaagtcacgctggccgacgccggcttcataaaacagtatggcgattgtcttggagacatcgccgcccgcgacctcatttgcgcacaaaagttcaatgggctcaccgtgttgccaccactgctcacagatgagatgatcgcacagtacacgagcgcccttcttgccggcactatcacgtctggttggacgttcggtgccggagccgctctgcaaattccctttgcaatgcaaatggcctatagatttaatggaattggcgtaacacagaacgtgttgtacgagaaccagaagctcattgccaaccagttcaattccgctattggcaaaatacaagactctctcagctcaactgctagcgcactgggaaaattgcaagacgtagtcaatcaaaatgcccaagccctcaatactctcgtcaaacagttgtcttccaactttggggctatcagtagtgtactcaatgacattctttcaagactggacccgcccgaggcggaagtccaaattgatcgtctgataactggaaggttgcaaagccttcagacctacgttacgcaacaacttattagggctgccgaaataagggcatccgctaatctggcagctacaaagatgtctgaatgtgttttgggacagagcaaacgggttgacttctgcggtaaaggttaccatctcatgtcttttccacaaagcgcaccgcacggagtcgtcttcctgcatgtaacatacgtcccagcccaagaaaagaattttaccacagccccagccatctgccacgacggcaaggcgcatttcccaagggaaggcgtgtttgtatccaacgggacgcattggtttgtcactcaaaggaacttttacgaaccccaaattattaccactgataacaccttcgtttctgggaactgtgatgtcgtgattgggatagtaaacaacacggtatatgatccactgcaaccagaactggattccttcaaagaagagctggacaaatacttcaagaatcatactagtcctgacgtcgacctgggcgatatcagtggaatcaacgctagcgtcgtaaacattcaaaaggagatcgatagacttaacgaggtcgccaagaatctcaatgaaagcctcatcgatttgcaagaactcggaaaatatgagcaaagcggatcagggtacattccggaagcccccagggacggacaggcatatgtccgcaaggacggagaatgggttcttcttagcacttttctggggtaatga (SEQ ID NO: 104)gccaccauggauuggaccuggauacuuuuccucguggccgcagcaacaagaguccacuccucucagugcguuaaccugacuacuagaacccaauugcccccggcauacacaaacucuuucacccggggugucuacuaucccgacaaaguguuuagaaguagcgugcugcacagcacccaagaucucuuucugccauucuucucaaacgucaccugguuucacgccauccauguaagcgggaccaacggcacaaagcguuuugauaacccuguuuugccauucaaugauggcguguauuuugcuuccacugagaaaagcaacaucauuagaggguggauauuuggcacaacgcuugacuccaagacgcagagucuuuugauaguaaacaacgcaacuaauguggucauuaaagucugugaauuucaauuuugcaaugaccccuuccuuggagucuauuaccacaagaacaacaaaagcuggauggaaagcgaauuuagggucuacagcucugccaauaacugcacauucgaauacgucagccaaccauucuugauggaccuggaaggcaagcaaggaaacuuuaagaaucugagggaauuuguguuuaagaauaucgacggauauuuuaagaucuauuccaagcauacucccauuaaucucguucgugaccuuccucaggguuucucugcauuggaaccccucguagauuugcccauugggauuaauaucacuagauuccagacgcugcuugcacuccaucgaucuuaucugaccccuggugacuccucuuccggguggacggcgggugcugcagccuacuacguuggcuauuugcaaccuaggaccuuucuguugaaguauaaugagaaugggacuauuacugaugccguugauugcgcccucgauccgcugucagaaacaaagugcacccugaagagcuucacaguagaaaagggaaucuaucaaaccucaaauuuccgcguucaaccaacugaaucaaucgugcguuuuccuaacaucacaaaucuguguccguuuggagaaguauuuaaugcgacgcguuucgcaagcgucuacgcguggaaucgcaaacguaucucuaauugcguagcagauuauucugugcuguacaauagcgcaucuuucucaacguuuaagugcuacggcguuaaugggaccaagcugaaugaucucuguuucacuaauguguacgcagacaguuuuguaauuagaggagacgagguuaggcaaauagcaccgggucaaacuggcaaaaucgccgacuauaacuacaagcucccugaugacuucacgggcugcguaauugcuuggaacucuaauaaccuggacucuaaagucggcgggaauuauaauuaucucuaucgguuguuucgaaaauccaaucucaaacccuuugagcgggacaucaauacuacaauuuaucaagcugguaguacuccuugcaaugggguagaaggcuucaauuguuauuucccccuucaaucuuacggauuucaacccacgaacggcguaggguaccagcccuaucgaguggugguacugucauucgaacuuaaucacgccccagcaacagucugcgggccuaagaaaagcacgaaucuugucaagaauaaguguguaaauuucaacuucaauggucuuacaggcacgggagugcucacugagucuaauaagaaauuucuuccuuuccaacaauucggucgugauauugccgauacuacugaugcaguccgagauccacaaacucucgaaauccucgauauuacuccuuguaguuuuggcggcgucuccgugaucaccccagggaccaacacuaguaaccaaguggcggugcucuaccaagauguuaacugcacagaagucccgguagcgauccaugccgaccagcucacucccacauggcguguuuacagcacagggucaaacguuuuccagacccgugccggaugucuuauaggagccgaacacguaaauaacaguuaugaaugcgauaucccaauuggugcagguaucugugcgucauaucaaacccaaacuaauucuccgggguccgccucaagcguugccucacaaucaauaaucgccuacacaaugucccucggugccgaaaauucagucgcuuacucuaacaauagcauugcuaucccuaccaacuucacuauuucuguuaccacggaaauuuugccuguauccaugaccaaaacaucuguugauugcacgauguacaucugcggggauucuaccgaauguucuaaccugcuucugcaauacggcuccuucugcacccaauugaaccgcgcacugacugggauugcuguggaacaagacaagaauacucaagaaguauuugcccaggucaaacagauuuacaaaacucccccaauuaaagauuucggcgguuucaauuuuagucaaauucugccagauccaaguaagccauccaaacgcucauuuauugaggaccugcucuuuaauaaagucacgcuggccgacgccggcuucauaaaacaguauggcgauugucuuggagacaucgccgcccgcgaccucauuugcgcacaaaaguucaaugggcucaccguguugccaccacugcucacagaugagaugaucgcacaguacacgagcgcccuucuugccggcacuaucacgucugguuggacguucggugccggagccgcucugcaaauucccuuugcaaugcaaauggccuauagauuuaauggaauuggcguaacacagaacguguuguacgagaaccagaagcucauugccaaccaguucaauuccgcuauuggcaaaauacaagacucucucagcucaacugcuagcgcacugggaaaauugcaagacguagucaaucaaaaugcccaagcccucaauacucucgucaaacaguugucuuccaacuuuggggcuaucaguaguguacucaaugacauucuuucaagacuggacccgcccgaggcggaaguccaaauugaucgucugauaacuggaagguugcaaagccuucagaccuacguuacgcaacaacuuauuagggcugccgaaauaagggcauccgcuaaucuggcagcuacaaagaugucugaauguguuuugggacagagcaaacggguugacuucugcgguaaagguuaccaucucaugucuuuuccacaaagcgcaccgcacggagucgucuuccugcauguaacauacgucccagcccaagaaaagaauuuuaccacagccccagccaucugccacgacggcaaggcgcauuucccaagggaaggcguguuuguauccaacgggacgcauugguuugucacucaaaggaacuuuuacgaaccccaaauuauuaccacugauaacaccuucguuucugggaacugugaugucgugauugggauaguaaacaacacgguauaugauccacugcaaccagaacuggauuccuucaaagaagagcuggacaaauacuucaagaaucauacuaguccugacgucgaccugggcgauaucaguggaaucaacgcuagcgucguaaacauucaaaaggagaucgauagacuuaacgaggucgccaagaaucucaaugaaagccucaucgauuugcaagaacucggaaaauaugagcaaagcggaucaggguacauuccggaagcccccagggacggacaggcauauguccgcaaggacggagaauggguucuucuuagcacuuuucugggguaauga (SEQ ID NO: 105)MDWTWILFLVAAATRVHSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGV

TKLNDLCFTNVY ADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDI

T

IYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRV VVLSFEL

HAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPGSASSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 106)WuS_DownDS3_D2P_Furin_pVaxgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcaatgttcgttttcctcgtgctcttgcctttggtttcttctcagtgcgtaaacctcacgactcgaacccaactgcccccagcttatacaaattcctttacgcggggcgtctattacccggataaggttttcagatccagcgtgctgcatagtacacaagatctctttcttcctttcttctcaaatgtaacctggtttcacgctattcatgtatccggcaccaatggaactaaaagatttgataacccggtgttgcccttcaatgatggtgtgtatttcgcttccacggaaaagtcaaacatcatcagagggtggatattcggcacaacattggattccaagtgccagtcactcctcatagtgaacaatgctactaacgtggttataaaggtctgcgaatttcaattttgtaatgatcctttcctcggtgtttactatcacaagaacaataagtcctggatggaatcagaattccgtgtatacagttctgcgaacaattgcacattcgaatatgtgtcccaaccctttctcatggatctggaagggaagcagggtaactttaagaatctgagagaattcgtgttcaagaacattgactgctattttaaaatctatagcaaacacacccctataaacttggtacgggatttgcctcaaggattctcagcactcgaacccttggtcgatttgccaatcggcatcaatatcacccggtttcagacactcctggctcttcaccgctcctacttgacacctggtgattcctcatctggttggaccgccggcgcagcggcatactatgtcggctatcttcaaccaagaaccttcttgctgaaatataatgagaacggaactataactgatgccgttgattgtgcccttgatccacttagcgaaacaaagtgcactctgaagtccttcacagttgaaaaggggatctaccaaacatccaacttccgggtacaacctactgagtccatagtgcgatttcctaacattaccaatctgtgcccatttggagaagtattcaacgcaactaggttcgcgtccgtttacgcgtggaacaggaaaaggatttccaattgcgtcgccgactatagcgttctctataacagcgcctcatttagcacgtttaagtgttacggggttagtccgaccaaactcaatgacttgtgttttaccaatgtctatgcagactcctttgttattagaggcgacgaggtcagacaaattgcccccggacagacaggtaagattgcagattataattataaactgccggacgacttcacggggtgtgttattgcatggaactccaataacctggactctaaagtaggcgggaactataactatctgtatcgcctgtttcgcaaatctaacctgaaacccttctgcagggacatatgtactgaaatatatcaagctggcagcacaccttgtaatggcgtcgagggattcaattgttacttcccacttcaatcttacggttttcagcctactaacggcgtagggtatcaaccctatagagttgtagtgctctctttcgaattgctccatgcccccgcgactgtttgtggacctaagaagtccacgaacctggtaaagaacaagtgtgttaattttaattttaatggactgaccgggactggagtgctgactgaaagtaacaagaaatttctgcctttccaacaatttggccgcgatatcgctgataccaccgacgccgtcagagatccgcagactctcgaaatcctggacatcacgccctgctcattcggcggggttagcgttattactccaggcactaacactagcaatcaagttgcagttctgtaccaggatgtgaactgtaccgaagtccccgtcgccattcatgccgatcagctgaccccgacttggcgggtatattcaaccggcagcaatgtctttcaaacaagggcgggttgtctcatcggagcggagcatgtaaataatagttatgaatgcgacatccccattggcgcggggatctgtgcttcatatcaaactcaaaccaattccccacggcggagacgatcagtagccagtcaatcaataattgcgtatacgatgagtcttggggcagaaaatagcgtggcttattctaataatagcatcgctatacctacaaattttacaatcagtgtaactaccgaaatccttcctgtcagcatgaccaaaactagcgtagattgcacgatgtatatttgcggagactcaactgagtgcagtaacctgttgttgcaatacggaagtttctgtacccagctgaaccgcgctcttacgggcattgcagtagaacaagataagaatacccaagaagtgtttgcccaggtgaaacaaatctacaagactcccccgattaaagactttggcgggttcaacttcagccagatattgcccgacccgtctcgtcgtagacggtcctttattgaagacctgctcttcaacaaggtcacactggctgatgcaggttttattaagcaatacggcgactgtcttggcgacatcgccgctagggaccttatatgtgctcagaaattcaatggtctgacagttctgccacccttgctcactgacgaaatgatcgctcaatatacaagcgccttgctggctgggactattacttccggatggacattcggggcgggtgccgccttgcaaattccttttgcaatgcaaatggcataccgtttcaacggaatcggcgtaacccagaatgtgctctatgaaaaccagaaattgatagcaaatcaatttaactcagccataggaaagattcaagactctctcagctcaaccgcgagtgctctcggcaagctccaagacgtagtaaatcaaaatgcacaagctttgaacactttggtaaagcaattgtcttccaacttcggggcgatctcatctggccctaacgacatcctgtcccggttgcccaaagtggaagccgaggtgcagatcgaccgcctcatcaccggccgacttcaatcactccaaacctacgtgactcaacaactgatccgggcagccgagataagggcgagtgcaaacttggcagctacgaaaatgtcagaatgtgttctcggccagagtaaacgggtagacttttgtgggaaaggttatcacttgatgtctttccctcaaagcgctcctcacggcgtcgtcttcttgcatgtgacttacgtgccagctcaagaaaagaacttcaccaccgcccctgctatatgccatgacggtaaagctcacttcccccgagagggcgtgttcgttagtaatggaacccattggtttgtgactcaacgaaacttttatgaacctcaaataattaccacggataacacttttgttagtggtaattgtgacgtggtgatcggcattgtgaataacacagtctacgatcctctgcaaccagaactggacagctttaaagaggaacttgacaaatatttcaagaaccatacaagccccgacgtcgacctgggcgacatcagtggaatcaatgcgtccgtagtcaatatccagaaggagattgatcggcttaatgaagtcgctaagaatttgaatgaaagtcttatagatctgcaagaactcgggaagtacgagcaatatattaaatggccttggtccggacgtagaaggcgcaggcggggctcaggcggttcagggtcagggtatattcccgaggcgccacgcgatgggcaagcgtacgtgcgtaaagatggcgaatgggtgttgctttccacattcttggggtgataa (SEQ ID NO: 107)gccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaauguucguuuuccucgugcucuugccuuugguuucuucucagugcguaaaccucacgacucgaacccaacugcccccagcuuauacaaauuccuuuacgcggggcgucuauuacccggauaagguuuucagauccagcgugcugcauaguacacaagaucucuuucuuccuuucuucucaaauguaaccugguuucacgcuauucauguauccggcaccaauggaacuaaaagauuugauaacccgguguugcccuucaaugaugguguguauuucgcuuccacggaaaagucaaacaucaucagaggguggauauucggcacaacauuggauuccaagugccagucacuccucauagugaacaaugcuacuaacgugguuauaaaggucugcgaauuucaauuuuguaaugauccuuuccucgguguuuacuaucacaagaacaauaaguccuggauggaaucagaauuccguguauacaguucugcgaacaauugcacauucgaauaugugucccaacccuuucucauggaucuggaagggaagcaggguaacuuuaagaaucugagagaauucguguucaagaacauugacugcuauuuuaaaaucuauagcaaacacaccccuauaaacuugguacgggauuugccucaaggauucucagcacucgaacccuuggucgauuugccaaucggcaucaauaucacccgguuucagacacuccuggcucuucaccgcuccuacuugacaccuggugauuccucaucugguuggaccgccggcgcagcggcauacuaugucggcuaucuucaaccaagaaccuucuugcugaaauauaaugagaacggaacuauaacugaugccguugauugugcccuugauccacuuagcgaaacaaagugcacucugaaguccuucacaguugaaaaggggaucuaccaaacauccaacuuccggguacaaccuacugaguccauagugcgauuuccuaacauuaccaaucugugcccauuuggagaaguauucaacgcaacuagguucgcguccguuuacgcguggaacaggaaaaggauuuccaauugcgucgccgacuauagcguucucuauaacagcgccucauuuagcacguuuaaguguuacgggguuaguccgaccaaacucaaugacuuguguuuuaccaaugucuaugcagacuccuuuguuauuagaggcgacgaggucagacaaauugcccccggacagacagguaagauugcagauuauaauuauaaacugccggacgacuucacgggguguguuauugcauggaacuccaauaaccuggacucuaaaguaggcgggaacuauaacuaucuguaucgccuguuucgcaaaucuaaccugaaacccuucugcagggacauauguacugaaauauaucaagcuggcagcacaccuuguaauggcgucgagggauucaauuguuacuucccacuucaaucuuacgguuuucagccuacuaacggcguaggguaucaacccuauagaguuguagugcucucuuucgaauugcuccaugcccccgcgacuguuuguggaccuaagaaguccacgaaccugguaaagaacaaguguguuaauuuuaauuuuaauggacugaccgggacuggagugcugacugaaaguaacaagaaauuucugccuuuccaacaauuuggccgcgauaucgcugauaccaccgacgccgucagagauccgcagacucucgaaauccuggacaucacgcccugcucauucggcgggguuagcguuauuacuccaggcacuaacacuagcaaucaaguugcaguucuguaccaggaugugaacuguaccgaaguccccgucgccauucaugccgaucagcugaccccgacuuggcggguauauucaaccggcagcaaugucuuucaaacaagggcggguugucucaucggagcggagcauguaaauaauaguuaugaaugcgacauccccauuggcgcggggaucugugcuucauaucaaacucaaaccaauuccccacggcggagacgaucaguagccagucaaucaauaauugcguauacgaugagucuuggggcagaaaauagcguggcuuauucuaauaauagcaucgcuauaccuacaaauuuuacaaucaguguaacuaccgaaauccuuccugucagcaugaccaaaacuagcguagauugcacgauguauauuugcggagacucaacugagugcaguaaccuguuguugcaauacggaaguuucuguacccagcugaaccgcgcucuuacgggcauugcaguagaacaagauaagaauacccaagaaguguuugcccaggugaaacaaaucuacaagacucccccgauuaaagacuuuggcggguucaacuucagccagauauugcccgacccgucucgucguagacgguccuuuauugaagaccugcucuucaacaaggucacacuggcugaugcagguuuuauuaagcaauacggcgacugucuuggcgacaucgccgcuagggaccuuauaugugcucagaaauucaauggucugacaguucugccacccuugcucacugacgaaaugaucgcucaauauacaagcgccuugcuggcugggacuauuacuuccggauggacauucggggcgggugccgccuugcaaauuccuuuugcaaugcaaauggcauaccguuucaacggaaucggcguaacccagaaugugcucuaugaaaaccagaaauugauagcaaaucaauuuaacucagccauaggaaagauucaagacucucucagcucaaccgcgagugcucucggcaagcuccaagacguaguaaaucaaaaugcacaagcuuugaacacuuugguaaagcaauugucuuccaacuucggggcgaucucaucuggcccuaacgacauccugucccgguugcccaaaguggaagccgaggugcagaucgaccgccucaucaccggccgacuucaaucacuccaaaccuacgugacucaacaacugauccgggcagccgagauaagggcgagugcaaacuuggcagcuacgaaaaugucagaauguguucucggccagaguaaacggguagacuuuugugggaaagguuaucacuugaugucuuucccucaaagcgcuccucacggcgucgucuucuugcaugugacuuacgugccagcucaagaaaagaacuucaccaccgccccugcuauaugccaugacgguaaagcucacuucccccgagagggcguguucguuaguaauggaacccauugguuugugacucaacgaaacuuuuaugaaccucaaauaauuaccacggauaacacuuuuguuagugguaauugugacguggugaucggcauugugaauaacacagucuacgauccucugcaaccagaacuggacagcuuuaaagaggaacuugacaaauauuucaagaaccauacaagccccgacgucgaccugggcgacaucaguggaaucaaugcguccguagucaauauccagaaggagauugaucggcuuaaugaagucgcuaagaauuugaaugaaagucuuauagaucugcaagaacucgggaaguacgagcaauauauuaaauggccuugguccggacguagaaggcgcaggcggggcucaggcgguucagggucaggguauauucccgaggcgccacgcgaugggcaagcguacgugcguaaagauggcgaauggguguugcuuuccacauucuuggggugauaa (SEQ ID NO: 108)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKCQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNID

YFKIYSKHTPIN LVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF

RDI

TEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICA SYQTQTNSPRR

RSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPS

RSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVK QLSSNFGAISS

NDILSRL

KVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 109) WuS_DownDS3_D2P_F_NoTriCle_pVaxgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcaatgttcgtgttcctcgtgctcctgcctctcgttagcagccaatgtgttaatctcaccaccagaacacagctcccacccgcgtatactaactcttttacgaggggagtttattatcccgataaggttttccggtctagcgtactccactccacccaagatctgttcctgcctttctttagcaacgtgacgtggtttcatgcaatccacgtgagtggcaccaatggaaccaagcggttcgataatcctgtgttgccgtttaacgatggcgtgtattttgcctcaactgaaaagtctaacataatacgcggctggatcttcgggaccacattggatagtaagtgtcaatctctgcttatcgtgaacaacgctactaacgtcgttataaaggtctgtgagttccaattctgcaacgacccattcctgggtgtgtattaccacaagaataataaatcttggatggagtctgagtttcgcgtatactcttctgctaacaactgcacctttgaatatgtaagtcaaccattcctcatggatctggaaggaaaacaaggcaactttaagaacttgcgggaatttgtcttcaagaacatcgactgttattttaaaatttactcaaaacacaccccgattaatctggtccgcgatttgccccaagggttctctgcattggaaccccttgtggacttgcctataggaattaatatcacccgctttcaaactcttctggcgctgcaccgtagctacctgacaccaggagatagctctagtggctggactgctggagccgcggcatattatgtggggtatctgcagccacgtacatttctcctcaaatataatgaaaatggtacaataacggatgcagtcgactgcgcattggaccctctgagtgaaacaaaatgcactctcaagagcttcactgttgaaaagggcatataccaaacatctaattttagagtccaacccactgaatccattgtccgatttcctaatattacaaacctctgcccatttggagaagtgttcaacgccactaggtttgcatccgtgtacgcatggaacagaaaacgaatttctaattgtgtggcagactatagcgtgctgtataactcagcaagctttagcacatttaagtgttatggagttagcccaaccaaattgaatgatctttgtttcacgaacgtgtacgccgatagcttcgttattcgaggggacgaggtgaggcaaatcgctccaggtcaaaccggtaaaatcgccgattacaattataaacttcctgatgacttcactggctgtgtcatagcatggaactctaataatctcgacagcaaggtcggtgggaactataactatctttatcgactctttagaaagagtaatctcaaaccattttgcagagacatttgtacagagatttatcaggcagggagcacaccatgtaatggggtcgagggcttcaactgttacttccccctgcaatcttatgggttccagccgaccaatggagtgggctaccaaccttatcgcgtggtggtcctgtcttttgaactgcttcatgctccagccaccgtatgcggccctaagaagtctacaaatttggtcaagaacaagtgcgtcaattttaacttcaatggtctgactggaaccggtgtcctcacagaatctaacaagaaatttctgccatttcaacaatttggaagagatatcgcggatactacggatgctgttagggacccccaaacacttgaaattctcgacattacaccctgttcctttggcggggtcagtgtcattaccccgggtacaaatactagtaaccaagtcgcagtactgtatcaagatgttaattgtaccgaagtgccggtagcaatacacgctgatcaacttacaccaacatggcgagtgtattctacggggagtaatgtcttccaaacgcgggccgggtgtctgattggcgcggaacacgtaaacaactcctacgaatgtgatattccaataggcgcaggcatatgtgcgagctatcaaacacaaactaactcccctagacggcgtcggagtgtggctagtcaatcaatcattgcctatacaatgtctctgggagcagaaaacagcgtggcatattccaataattccatcgctatacctaccaactttaccatcagcgtcactactgagattcttcccgtctccatgacgaaaacttccgttgattgtactatgtacatctgcggagacagcaccgaatgcagtaaccttctcttgcaatatggcagcttttgtactcagctcaacagagctctcacaggtattgccgtcgaacaagataagaacacccaagaggtgttcgcccaggtgaaacagatatataagaccccacccatcaaggatttcggcgggtttaattttagtcaaatcctgcccgatccctcacggcgtcgcaggtcctttattgaagatcttctgttcaataaggtcacactcgctgacgcaggctttatcaagcagtatggagattgtctgggcgatatagctgcgagggacttgatctgcgcacaaaagttcaacggccttacagtgctgcccccgttgctgacagatgagatgattgcgcaatacacttccgcgcttctcgcagggaccatcacgagcggctggacgttcggcgctggcgccgctctgcaaatcccgtttgcaatgcaaatggcctataggtttaatggtatcggtgtaacgcaaaacgtactttatgaaaaccagaaactgatcgctaaccaattcaattccgctattggcaaaattcaagacagcctcagcagcacggctagtgcactgggtaaactccaagacgtggtgaaccaaaatgcccaagcattgaatacacttgtcaagcaacttagttccaacttcggtgcaatttcaagtggtccaaatgacatacttagcaggctgcctaaagtagaagccgaagtgcaaatcgatagacttatcaccggccgcctgcaatcccttcaaacatacgtgactcagcagcttatcagggctgctgagattcgagcaagtgcgaacctggccgccaccaaaatgagtgagtgcgtccttgggcaatccaagcgcgttgacttttgtggtaaggggtatcatctcatgagcttcccccaatccgcccctcacggagtagtgtttctccatgtgacgtatgttcctgcacaagagaagaacttcacaacggctccggctatatgtcatgacggaaaagcgcactttcctcgcgaaggagtgtttgtgtcaaatggaacgcactggttcgtgacgcaaaggaatttctacgagcctcaaatcatcactacagataatacttttgtctctgggaattgcgacgtggtcattggaatcgtcaacaatacggtttacgatcccctgcaaccagaactggattcattcaaagaagaactcgacaagtacttcaagaatcataccagtcctgatgtggatctgggcgatatcagtgggatcaatgcaagcgttgtcaacattcaaaaggaaatagaccgcctcaacgaagtcgcaaagaatctcaatgaaagccttattgatcttcaagagctcggaaaatatgagcaatatattaagtggccttggtccggcggctcaggcggaagtggctcaggatatattcctgaggctccccgagatggacaagcatacgtgagaaaagatggggagtgggtgttgctgagtacgttccttggatgataa (SEQ ID NO: 110)gccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaauguucguguuccucgugcuccugccucucguuagcagccaauguguuaaucucaccaccagaacacagcucccacccgcguauacuaacucuuuuacgaggggaguuuauuaucccgauaagguuuuccggucuagcguacuccacuccacccaagaucuguuccugccuuucuuuagcaacgugacgugguuucaugcaauccacgugaguggcaccaauggaaccaagcgguucgauaauccuguguugccguuuaacgauggcguguauuuugccucaacugaaaagucuaacauaauacgcggcuggaucuucgggaccacauuggauaguaagugucaaucucugcuuaucgugaacaacgcuacuaacgucguuauaaaggucugugaguuccaauucugcaacgacccauuccuggguguguauuaccacaagaauaauaaaucuuggauggagucugaguuucgcguauacucuucugcuaacaacugcaccuuugaauauguaagucaaccauuccucauggaucuggaaggaaaacaaggcaacuuuaagaacuugcgggaauuugucuucaagaacaucgacuguuauuuuaaaauuuacucaaaacacaccccgauuaaucugguccgcgauuugccccaaggguucucugcauuggaaccccuuguggacuugccuauaggaauuaauaucacccgcuuucaaacucuucuggcgcugcaccguagcuaccugacaccaggagauagcucuaguggcuggacugcuggagccgcggcauauuaugugggguaucugcagccacguacauuucuccucaaauauaaugaaaaugguacaauaacggaugcagucgacugcgcauuggacccucugagugaaacaaaaugcacucucaagagcuucacuguugaaaagggcauauaccaaacaucuaauuuuagaguccaacccacugaauccauuguccgauuuccuaauauuacaaaccucugcccauuuggagaaguguucaacgccacuagguuugcauccguguacgcauggaacagaaaacgaauuucuaauuguguggcagacuauagcgugcuguauaacucagcaagcuuuagcacauuuaaguguuauggaguuagcccaaccaaauugaaugaucuuuguuucacgaacguguacgccgauagcuucguuauucgaggggacgaggugaggcaaaucgcuccaggucaaaccgguaaaaucgccgauuacaauuauaaacuuccugaugacuucacuggcugugucauagcauggaacucuaauaaucucgacagcaaggucggugggaacuauaacuaucuuuaucgacucuuuagaaagaguaaucucaaaccauuuugcagagacauuuguacagagauuuaucaggcagggagcacaccauguaauggggucgagggcuucaacuguuacuucccccugcaaucuuauggguuccagccgaccaauggagugggcuaccaaccuuaucgcguggugguccugucuuuugaacugcuucaugcuccagccaccguaugcggcccuaagaagucuacaaauuuggucaagaacaagugcgucaauuuuaacuucaauggucugacuggaaccgguguccucacagaaucuaacaagaaauuucugccauuucaacaauuuggaagagauaucgcggauacuacggaugcuguuagggacccccaaacacuugaaauucucgacauuacacccuguuccuuuggcggggucagugucauuaccccggguacaaauacuaguaaccaagucgcaguacuguaucaagauguuaauuguaccgaagugccgguagcaauacacgcugaucaacuuacaccaacauggcgaguguauucuacggggaguaaugucuuccaaacgcgggccgggugucugauuggcgcggaacacguaaacaacuccuacgaaugugauauuccaauaggcgcaggcauaugugcgagcuaucaaacacaaacuaacuccccuagacggcgucggaguguggcuagucaaucaaucauugccuauacaaugucucugggagcagaaaacagcguggcauauuccaauaauuccaucgcuauaccuaccaacuuuaccaucagcgucacuacugagauucuucccgucuccaugacgaaaacuuccguugauuguacuauguacaucugcggagacagcaccgaaugcaguaaccuucucuugcaauauggcagcuuuuguacucagcucaacagagcucucacagguauugccgucgaacaagauaagaacacccaagagguguucgcccaggugaaacagauauauaagaccccacccaucaaggauuucggcggguuuaauuuuagucaaauccugcccgaucccucacggcgucgcagguccuuuauugaagaucuucuguucaauaaggucacacucgcugacgcaggcuuuaucaagcaguauggagauugucugggcgauauagcugcgagggacuugaucugcgcacaaaaguucaacggccuuacagugcugcccccguugcugacagaugagaugauugcgcaauacacuuccgcgcuucucgcagggaccaucacgagcggcuggacguucggcgcuggcgccgcucugcaaaucccguuugcaaugcaaauggccuauagguuuaaugguaucgguguaacgcaaaacguacuuuaugaaaaccagaaacugaucgcuaaccaauucaauuccgcuauuggcaaaauucaagacagccucagcagcacggcuagugcacuggguaaacuccaagacguggugaaccaaaaugcccaagcauugaauacacuugucaagcaacuuaguuccaacuucggugcaauuucaagugguccaaaugacauacuuagcaggcugccuaaaguagaagccgaagugcaaaucgauagacuuaucaccggccgccugcaaucccuucaaacauacgugacucagcagcuuaucagggcugcugagauucgagcaagugcgaaccuggccgccaccaaaaugagugagugcguccuugggcaauccaagcgcguugacuuuugugguaagggguaucaucucaugagcuucccccaauccgccccucacggaguaguguuucuccaugugacguauguuccugcacaagagaagaacuucacaacggcuccggcuauaugucaugacggaaaagcgcacuuuccucgcgaaggaguguuugugucaaauggaacgcacugguucgugacgcaaaggaauuucuacgagccucaaaucaucacuacagauaauacuuuugucucugggaauugcgacguggucauuggaaucgucaacaauacgguuuacgauccccugcaaccagaacuggauucauucaaagaagaacucgacaaguacuucaagaaucauaccaguccugauguggaucugggcgauaucagugggaucaaugcaagcguugucaacauucaaaaggaaauagaccgccucaacgaagucgcaaagaaucucaaugaaagccuuauugaucuucaagagcucggaaaauaugagcaauauauuaaguggccuugguccggcggcucaggcggaaguggcucaggauauauuccugaggcuccccgagauggacaagcauacgugagaaaagauggggaguggguguugcugaguacguuccuuggaugauaa (SEQ ID NO: 111)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSK

QSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNID

YFKIYSKHTPIN LVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF

RDI

TEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICA SYQTQTNSPRR

RSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPS

RSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVK QLSSNFGAISS

NDILSRL

KVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG**(SEQ ID NO: 112) WuS_DownDS2_2P_pVaxgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcaatgttcgtgttcttggtgctgctgcctcttgtctcatcacagtgcgttaatctgaccacccgtacacaactcccacccgcatacacaaatagctttacacgcggagtgtattaccccgataaagtctttcggagctcagtgctccattctactcaagatcttttcctgccgttctttagtaacgttacttggtttcatgcaatacatgtgtctggcacaaacggaaccaaacgttttgataatccggtgttgccatttaatgatggtgtatattttgcttccacggaaaagtcaaacatcatccgtgggtggatctttggcaccactcttgatagcaaatgtcaaagccttctgattgttaataacgctacaaacgtcgtaattaaagtgtgtgaattccagttctgtaatgaccccttcctcggagtatattaccacaagaataacaaatcttggatggagagcgaatttagagtttacagttcagccaataactgtacatttgaatatgtcagtcagcctttcctcatggacctcgaaggtaaacaaggtaattttaagaacttgagagagttcgtgtttaagaacatcgatggctatttcaaaatttactctaagcacacaccaatcaacctggttcgagacctgccccagggtttctcagctttggaaccattggtggacctgccaatcggcattaacattaccagatttcaaactttgttggcactccaccggtcatatcttacccccggagacagttcctcaggctggacggcaggcgccgccgcgtactatgttgggtatctccaaccccgaaccttccttctcaaatacaatgaaaacgggacgattacagatgcagtcgattgcgccctggaccccttgtccgaaactaaatgcactctgaagagtttcacggtagagaagggaatctatcaaacgagcaattttcgagtccaaccaacggaatctattgtgcggtttcccaatatcacaaacctctgtccattcggagaagtctttaatgctaccaggtttgcgtctgtatatgcatggaaccgaaagaggatttccaattgcgtagcggactacagtgtcctttataacagcgcttcattttccacgtttaagtgttatggtgtttctccaacgaaactcaacgacctctgttttactaacgtttacgctgacagctttgttatacgtggggacgaagtcaggcaaattgctcctggacagactggaaagatcgctgattataattataaacttcctgacgatttcaccggctgcgttattgcatggaactccaacaatctggattcaaaagtgggtggaaattataattatctgtataggttgtttcggaagagcaatcttaagccctttgagcgggacatatgtaccgaaatttaccaagcaggctccaccccatgcaatggagtagaagggttcaattgctattttcctctgcaaagttatggctttcaacccaccaacggagttgggtatcaaccttacagggttgtcgtgctgagtttcgaattgctccacgcacccgctacagtatgtggccccaagaagtccactaatcttgttaagaataaatgcgtgaacttcaacttcaatggacttacaggtactggagtactcacggaatcaaacaagaaatttctcccatttcaacagtttggccgagatatagctgacaccacagatgctgttcgcgacccccagacgttggaaatacttgatatcactccctgcagcttcggcggcgtgagcgtgatcactccaggtactaatacgagcaatcaagttgccgttctgtaccaagatgtgaactgcaccgaggttccagtggcaattcacgccgaccaacttactcccacctggcgggtctattccaccggatcaaacgtcttccaaactcgcgctggttgccttatcggtgcagagcacgttaataattcctatgaatgtgacattcccataggagcaggcatctgtgcatcttatcaaacccagactaattcccctggttccgcttcctctgttgcatcccagtccataattgcctacactatgagtctcggggctgaaaattccgtggcctattctaataattcaatcgccatcccaaccaattttaccatatccgtaacgactgaaatacttcctgtcagtatgaccaagacctcagtggactgcaccatgtacatctgcggcgattctactgaatgttccaatctgcttttgcaatatggttcattctgcacccaactcaacagggctcttacagggatcgccgtcgaacaggataagaatacccaggaagtgttcgcccaagttaagcaaatttacaagacaccacccatcaaggacttcggcgggttcaacttcagccaaattctgcccgacccgtctaagccttctaagcgctctttcattgaggatcttttgttcaataaggttacgcttgccgatgcagggtttatcaaacagtatggcgactgtcttggggatatcgcagctagggatcttatttgtgcacagaaatttaatggcctgactgttcttccccctttgctcactgacgagatgattgcccagtacacttcagctctcctggccgggactataacttctggttggaccttcggagctggcgccgccctgcaaattccatttgcaatgcagatggcttatcgcttcaacggaattggggtgacccaaaatgttctctacgagaaccagaaactcattgcaaaccagttcaattctgcgatcgggaagatccaggattccctgtctagtacggctagtgccctcggtaagctccaagacgtcgtcaaccaaaacgcccaggccttgaacacccttgtcaaacaactgagctccaattttggggctattagcagtgtgctgaatgatatcctgtcccgccttgacccaccggaagcggaagtccaaattgatcgactgatcactgggcgtctccaatcccttcaaacttacgtgacccaacaactcatccgagcagctgagattagggctagcgctaaccttgctgctactaagatgtcagagtgtgtcctcggccagtctaagagagtggacttttgtgggaaagggtaccacttgatgtcattcccacaaagcgccccacacggcgtggtgtttctccacgtcacttacgttccagctcaggaaaagaactttaccaccgcccccgctatatgtcatgatgggaaggcccactttcctcgtgaaggtgtctttgtcagcaatggcacacactggtttgtgacccaacggaatttctatgagcctcagattattaccacggataacactttcgtatcagggaattgtgatgtggttatcggcatcgttaataatacagtgtatgacccactgcagccagagcttgacagcttcaaagaagagctcgataagtactttaagaatcatacaagtcctgacgttgatcttggggatattagtgggattaacgccagcgtcgtcaatattcagaaagagattgacaggttgaacgaagtagctaagaatcttaatgaaagcctgatagatttgcaagaacttggtaagtatgagcaggggtacatacccgaggctcctcgggatgggcaggcctatgtacgcaaagacggtgaatgggtattgctcagcacttttctcggctgataa (SEQ ID NO: 113)gccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaauguucguguucuuggugcugcugccucuugucucaucacagugcguuaaucugaccacccguacacaacucccacccgcauacacaaauagcuuuacacgcggaguguauuaccccgauaaagucuuucggagcucagugcuccauucuacucaagaucuuuuccugccguucuuuaguaacguuacuugguuucaugcaauacaugugucuggcacaaacggaaccaaacguuuugauaauccgguguugccauuuaaugaugguguauauuuugcuuccacggaaaagucaaacaucauccguggguggaucuuuggcaccacucuugauagcaaaugucaaagccuucugauuguuaauaacgcuacaaacgucguaauuaaagugugugaauuccaguucuguaaugaccccuuccucggaguauauuaccacaagaauaacaaaucuuggauggagagcgaauuuagaguuuacaguucagccaauaacuguacauuugaauaugucagucagccuuuccucauggaccucgaagguaaacaagguaauuuuaagaacuugagagaguucguguuuaagaacaucgauggcuauuucaaaauuuacucuaagcacacaccaaucaaccugguucgagaccugccccaggguuucucagcuuuggaaccauugguggaccugccaaucggcauuaacauuaccagauuucaaacuuuguuggcacuccaccggucauaucuuacccccggagacaguuccucaggcuggacggcaggcgccgccgcguacuauguuggguaucuccaaccccgaaccuuccuucucaaauacaaugaaaacgggacgauuacagaugcagucgauugcgcccuggaccccuuguccgaaacuaaaugcacucugaagaguuucacgguagagaagggaaucuaucaaacgagcaauuuucgaguccaaccaacggaaucuauugugcgguuucccaauaucacaaaccucuguccauucggagaagucuuuaaugcuaccagguuugcgucuguauaugcauggaaccgaaagaggauuuccaauugcguagcggacuacaguguccuuuauaacagcgcuucauuuuccacguuuaaguguuaugguguuucuccaacgaaacucaacgaccucuguuuuacuaacguuuacgcugacagcuuuguuauacguggggacgaagucaggcaaauugcuccuggacagacuggaaagaucgcugauuauaauuauaaacuuccugacgauuucaccggcugcguuauugcauggaacuccaacaaucuggauucaaaaguggguggaaauuauaauuaucuguauagguuguuucggaagagcaaucuuaagcccuuugagcgggacauauguaccgaaauuuaccaagcaggcuccaccccaugcaauggaguagaaggguucaauugcuauuuuccucugcaaaguuauggcuuucaacccaccaacggaguuggguaucaaccuuacaggguugucgugcugaguuucgaauugcuccacgcacccgcuacaguauguggccccaagaaguccacuaaucuuguuaagaauaaaugcgugaacuucaacuucaauggacuuacagguacuggaguacucacggaaucaaacaagaaauuucucccauuucaacaguuuggccgagauauagcugacaccacagaugcuguucgcgacccccagacguuggaaauacuugauaucacucccugcagcuucggcggcgugagcgugaucacuccagguacuaauacgagcaaucaaguugccguucuguaccaagaugugaacugcaccgagguuccaguggcaauucacgccgaccaacuuacucccaccuggcgggucuauuccaccggaucaaacgucuuccaaacucgcgcugguugccuuaucggugcagagcacguuaauaauuccuaugaaugugacauucccauaggagcaggcaucugugcaucuuaucaaacccagacuaauuccccugguuccgcuuccucuguugcaucccaguccauaauugccuacacuaugagucucggggcugaaaauuccguggccuauucuaauaauucaaucgccaucccaaccaauuuuaccauauccguaacgacugaaauacuuccugucaguaugaccaagaccucaguggacugcaccauguacaucugcggcgauucuacugaauguuccaaucugcuuuugcaauaugguucauucugcacccaacucaacagggcucuuacagggaucgccgucgaacaggauaagaauacccaggaaguguucgcccaaguuaagcaaauuuacaagacaccacccaucaaggacuucggcggguucaacuucagccaaauucugcccgacccgucuaagccuucuaagcgcucuuucauugaggaucuuuuguucaauaagguuacgcuugccgaugcaggguuuaucaaacaguauggcgacugucuuggggauaucgcagcuagggaucuuauuugugcacagaaauuuaauggccugacuguucuucccccuuugcucacugacgagaugauugcccaguacacuucagcucuccuggccgggacuauaacuucugguuggaccuucggagcuggcgccgcccugcaaauuccauuugcaaugcagauggcuuaucgcuucaacggaauuggggugacccaaaauguucucuacgagaaccagaaacucauugcaaaccaguucaauucugcgaucgggaagauccaggauucccugucuaguacggcuagugcccucgguaagcuccaagacgucgucaaccaaaacgcccaggccuugaacacccuugucaaacaacugagcuccaauuuuggggcuauuagcagugugcugaaugauauccugucccgccuugacccaccggaagcggaaguccaaauugaucgacugaucacugggcgucuccaaucccuucaaacuuacgugacccaacaacucauccgagcagcugagauuagggcuagcgcuaaccuugcugcuacuaagaugucagaguguguccucggccagucuaagagaguggacuuuugugggaaaggguaccacuugaugucauucccacaaagcgccccacacggcgugguguuucuccacgucacuuacguuccagcucaggaaaagaacuuuaccaccgcccccgcuauaugucaugaugggaaggcccacuuuccucgugaaggugucuuugucagcaauggcacacacugguuugugacccaacggaauuucuaugagccucagauuauuaccacggauaacacuuucguaucagggaauugugaugugguuaucggcaucguuaauaauacaguguaugacccacugcagccagagcuugacagcuucaaagaagagcucgauaaguacuuuaagaaucauacaaguccugacguugaucuuggggauauuagugggauuaacgccagcgucgucaauauucagaaagagauugacagguugaacgaaguagcuaagaaucuuaaugaaagccugauagauuugcaagaacuugguaaguaugagcagggguacauacccgaggcuccucgggaugggcaggccuauguacgcaaagacggugaauggguauugcucagcacuuuucucggcugauaa (SEQ ID NO: 114)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSK

QSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDI

TEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICA SYQTQTNSP

A

SVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLD

EAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 115)WuSDownDS1_2P_pVaxgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcaatgtttgttttccttgttctgctcccgcttgtgtcttcccagtgcgtgaacctgaccacccgaactcaactcccaccagcatacaccaactcatttacaagaggagtttattacccggacaaggtatttcgaagttcagttcttcacagcacccaagacctgtttctgccattcttcagtaatgtcacttggtttcacgcgatacatgtcagcggtacaaacgggacaaagcgattcgataacccagtactcccattcaacgacggagtgtattttgcatctacagagaaatccaacattatacgcgggtggatctttggaactactctggactccaagacacagagcctgctcattgtgaacaatgcaacgaatgtcgtcataaaagtctgtgaatttcaattttgcaacgatcctttcctcggagtctattaccataagaacaataagagttggatggagagtgagtttcgcgtctattcttccgcgaacaattgtacatttgaatatgtatcacaaccctttcttatggatttggaaggcaaacaaggtaacttcaagaacttgcgcgagttcgtgttcaagaacatagactgttattttaagatctatagtaagcatacgccaatcaatctggtgcgagatttgcctcagggcttttctgctcttgaacccttggttgatctgcccatcgggatcaacataaccagatttcaaacgttgctcgcactccaccgcagctatctcactcctggcgattcctcatctgggtggaccgccggagctgctgcttattacgtcggctatctccagccgcgtactttcctgctcaagtataatgagaatggcaccattaccgatgctgtggattgtgctcttgatccactctctgaaaccaaatgcactctcaagtcttttaccgtggaaaagggtatttatcagacatctaattttcgggtgcaacctactgagtcaattgtacggtttcctaacataactaacctttgtccatttggggaagtcttcaatgccacgcggttcgcatcagtctatgcatggaacagaaaacgtatctccaactgcgtcgccgattattccgtcctttacaatagcgctagcttttccacattcaaatgttatggcgtatcaccaaccaaacttaacgatctctgctttactaatgtctacgctgactctttcgttattcgaggtgacgaggtgcgccaaattgcgcctggtcaaaccggaaagattgccgattataactacaagctccccgacgactttacgggttgtgtgatcgcctggaatagcaataacctcgattctaaagttggcggtaattataactatctgtacagactctttaggaaaagtaatctcaagcccttttgcagggatatctcaaccgaaatctaccaagccggcagcactccttgcaatggtgtcgaggggtttaattgttatttcccactgcaatcttacggctttcaaccgactaatggagtcggttatcaaccctatagggtggtggtactctcctttgaacttttgcacgctccggcaacagtttgtggaccaaagaaaagtacgaaccttgttaagaataagtgtgttaatttcaattttaacggcctcactggaacaggtgtcctcacagaaagcaacaagaagtttctccctttccaacagtttggacgggatatcgccgacactactgacgccgtcagagatcctcaaactctcgaaatcttggatatcacaccatgttctttcggtggtgtctccgtcataacaccaggaactaacacctctaatcaagtggccgtgctctatcaggacgtcaattgcacagaagtgcctgtcgcaatccatgctgatcagctcactcccacctggcgtgtgtattccactggctctaatgtctttcagacacgggcaggttgccttattggggcagagcatgtgaacaattcctacgaatgcgatatacccattggggcaggcatttgcgccagctaccaaacccaaactaacagccccgggagtgccagcagcgtggcatctcagtccattattgcctatacgatgagcctgggtgctgaaaatagcgtggcttatagtaataactctatcgccatacccacaaacttcaccatttcagtgaccaccgaaatccttcctgtttctatgaccaaaacgtccgtcgattgtacaatgtacatttgcggcgatagcactgaatgttcaaacctgctcctgcaatacggctctttctgtactcagctcaaccgggcactcaccggcatagccgtcgaacaagacaagaatacccaggaagtctttgcgcaggtgaaacaaatctataagaccccaccaataaaagatttcggcggttttaatttcagccaaatcttgcctgatcccagcaagccatctaaacggtctttcattgaagatctcctgttcaacaaggttacgctggctgacgccgggtttattaagcaatatggcgattgccttggggacattgccgcacgagacctcatttgtgcccagaaattcaacgggctcaccgtattgcccccgctcctcacagacgaaatgatcgcccaatatacaagcgccctgcttgcgggaaccattacaagcggttggacctttggtgccggcgcagctctgcaaatacccttcgcaatgcaaatggcatatcggtttaatggaattggcgtaacccaaaacgtgctgtatgaaaaccagaaactgatcgcaaatcaattcaatagtgctataggaaagatccaagacagtctgtcttccactgctagcgcgctggggaagctccaagacgttgtgaaccaaaacgcgcaggccctgaataccctggtgaagcaactttcaagcaatttcggtgctatatcttctgtcctcaatgacattctctctcggctcgatcccccggaagccgaagttcagatagaccgtttgatcacaggccgcttgcaatccctgcaaacctacgttacacaacaactgattcgcgccgccgaaattcgggcatccgccaatctggccgcaaccaaaatgtccgagtgtgttctcggtcaatccaaacgcgtggatttctgcggaaaaggataccatttgatgtcatttccacaatcagctccacacggtgttgtattcctgcacgtgacctacgtgccagcccaggagaagaattttactactgcgcccgccatttgtcatgacgggaaggctcattttcctcgggaaggggttttcgtctcaaacggtacccattggttcgtgactcagaggaacttttatgaacctcaaatcataacgaccgataacacgtttgtaagtggcaattgcgacgtggtcatcgggattgtaaacaatactgtctatgaccctctccaaccagagcttgacagctttaaagaagagcttgataaatactttaagaaccatacctcaccagacgtcgatttgggagatatcagtggcattaatgcctctgtcgtcaatatccagaaagagattgaccgcttgaacgaagttgccaagaatcttaatgagtctctgattgacttgcaagaattgggaaaatatgaacaaggatatattccagaagcccctcgcgatgggcaagcatatgttcgaaaggatggggaatgggtgctgctcagcacctttctcggttgataa (SEQ ID NO: 116)gccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaauguuuguuuuccuuguucugcucccgcuugugucuucccagugcgugaaccugaccacccgaacucaacucccaccagcauacaccaacucauuuacaagaggaguuuauuacccggacaagguauuucgaaguucaguucuucacagcacccaagaccuguuucugccauucuucaguaaugucacuugguuucacgcgauacaugucagcgguacaaacgggacaaagcgauucgauaacccaguacucccauucaacgacggaguguauuuugcaucuacagagaaauccaacauuauacgcggguggaucuuuggaacuacucuggacuccaagacacagagccugcucauugugaacaaugcaacgaaugucgucauaaaagucugugaauuucaauuuugcaacgauccuuuccucggagucuauuaccauaagaacaauaagaguuggauggagagugaguuucgcgucuauucuuccgcgaacaauuguacauuugaauauguaucacaacccuuucuuauggauuuggaaggcaaacaagguaacuucaagaacuugcgcgaguucguguucaagaacauagacuguuauuuuaagaucuauaguaagcauacgccaaucaaucuggugcgagauuugccucagggcuuuucugcucuugaacccuugguugaucugcccaucgggaucaacauaaccagauuucaaacguugcucgcacuccaccgcagcuaucucacuccuggcgauuccucaucuggguggaccgccggagcugcugcuuauuacgucggcuaucuccagccgcguacuuuccugcucaaguauaaugagaauggcaccauuaccgaugcuguggauugugcucuugauccacucucugaaaccaaaugcacucucaagucuuuuaccguggaaaaggguauuuaucagacaucuaauuuucgggugcaaccuacugagucaauuguacgguuuccuaacauaacuaaccuuuguccauuuggggaagucuucaaugccacgcgguucgcaucagucuaugcauggaacagaaaacguaucuccaacugcgucgccgauuauuccguccuuuacaauagcgcuagcuuuuccacauucaaauguuauggcguaucaccaaccaaacuuaacgaucucugcuuuacuaaugucuacgcugacucuuucguuauucgaggugacgaggugcgccaaauugcgccuggucaaaccggaaagauugccgauuauaacuacaagcuccccgacgacuuuacggguugugugaucgccuggaauagcaauaaccucgauucuaaaguuggcgguaauuauaacuaucuguacagacucuuuaggaaaaguaaucucaagcccuuuugcagggauaucucaaccgaaaucuaccaagccggcagcacuccuugcaauggugucgagggguuuaauuguuauuucccacugcaaucuuacggcuuucaaccgacuaauggagucgguuaucaacccuauaggguggugguacucuccuuugaacuuuugcacgcuccggcaacaguuuguggaccaaagaaaaguacgaaccuuguuaagaauaaguguguuaauuucaauuuuaacggccucacuggaacagguguccucacagaaagcaacaagaaguuucucccuuuccaacaguuuggacgggauaucgccgacacuacugacgccgucagagauccucaaacucucgaaaucuuggauaucacaccauguucuuucgguggugucuccgucauaacaccaggaacuaacaccucuaaucaaguggccgugcucuaucaggacgucaauugcacagaagugccugucgcaauccaugcugaucagcucacucccaccuggcguguguauuccacuggcucuaaugucuuucagacacgggcagguugccuuauuggggcagagcaugugaacaauuccuacgaaugcgauauacccauuggggcaggcauuugcgccagcuaccaaacccaaacuaacagccccgggagugccagcagcguggcaucucaguccauuauugccuauacgaugagccugggugcugaaaauagcguggcuuauaguaauaacucuaucgccauacccacaaacuucaccauuucagugaccaccgaaauccuuccuguuucuaugaccaaaacguccgucgauuguacaauguacauuugcggcgauagcacugaauguucaaaccugcuccugcaauacggcucuuucuguacucagcucaaccgggcacucaccggcauagccgucgaacaagacaagaauacccaggaagucuuugcgcaggugaaacaaaucuauaagaccccaccaauaaaagauuucggcgguuuuaauuucagccaaaucuugccugaucccagcaagccaucuaaacggucuuucauugaagaucuccuguucaacaagguuacgcuggcugacgccggguuuauuaagcaauauggcgauugccuuggggacauugccgcacgagaccucauuugugcccagaaauucaacgggcucaccguauugcccccgcuccucacagacgaaaugaucgcccaauauacaagcgcccugcuugcgggaaccauuacaagcgguuggaccuuuggugccggcgcagcucugcaaauacccuucgcaaugcaaauggcauaucgguuuaauggaauuggcguaacccaaaacgugcuguaugaaaaccagaaacugaucgcaaaucaauucaauagugcuauaggaaagauccaagacagucugucuuccacugcuagcgcgcuggggaagcuccaagacguugugaaccaaaacgcgcaggcccugaauacccuggugaagcaacuuucaagcaauuucggugcuauaucuucuguccucaaugacauucucucucggcucgaucccccggaagccgaaguucagauagaccguuugaucacaggccgcuugcaaucccugcaaaccuacguuacacaacaacugauucgcgccgccgaaauucgggcauccgccaaucuggccgcaaccaaaauguccgaguguguucucggucaauccaaacgcguggauuucugcggaaaaggauaccauuugaugucauuuccacaaucagcuccacacgguguuguauuccugcacgugaccuacgugccagcccaggagaagaauuuuacuacugcgcccgccauuugucaugacgggaaggcucauuuuccucgggaagggguuuucgucucaaacgguacccauugguucgugacucagaggaacuuuuaugaaccucaaaucauaacgaccgauaacacguuuguaaguggcaauugcgacguggucaucgggauuguaaacaauacugucuaugacccucuccaaccagagcuugacagcuuuaaagaagagcuugauaaauacuuuaagaaccauaccucaccagacgucgauuugggagauaucaguggcauuaaugccucugucgucaauauccagaaagagauugaccgcuugaacgaaguugccaagaaucuuaaugagucucugauugacuugcaagaauugggaaaauaugaacaaggauauauuccagaagccccucgcgaugggcaagcauauguucgaaaggauggggaaugggugcugcucagcaccuuucucgguugauaa(SEQ ID NO: 117)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNID

YFKIYSKHTPIN LVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPF

RDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICAS YQTQTNSP

A

SVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLD

EAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 118) WuS_2P_pVaxgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcaatgtttgtgtttcttgtcctgttgccattggtgagctcccaatgtgtcaatctgaccacccggacacaattgccccctgcatatacaaattcattcacgagaggagtatactatcccgacaaagttttccggtcctcagtccttcattccactcaagatcttttccttccattcttttctaacgtaacctggttccatgcaattcatgtcagtgggaccaacggcacgaaacggtttgataatccagttcttccattcaatgacggagtatattttgcatcaactgagaaatctaatatcattagagggtggattttcggaacaactcttgactccaagacccaatccttgctcatcgttaacaatgctacaaatgtggttattaaggtctgtgagtttcaattctgtaacgacccctttctcggcgtatactaccataagaataataagtcttggatggagtctgaatttcgtgtctactcatcagcgaacaattgtacatttgaatatgtgtcccaaccattcctgatggatctcgaaggaaagcagggcaattttaagaaccttcgggagttcgtctttaagaatatcgatggatactttaaaatatatagtaaacacacaccaatcaatctggtccgagatctcccccagggttttagtgctctggagccgctggtggatttgcccatcggtatcaatattacgcgcttccaaacattgctcgccctgcatcggagttaccttacgcctggcgacagtagcagcggatggaccgctggagccgccgcctactatgtcgggtaccttcaaccacgcacttttctcctgaaatacaacgaaaatgggacaattacagacgctgttgattgcgcactcgatcccctgtcagaaacaaaatgtacacttaaatcttttacggtcgagaaagggatttaccagacatctaacttccgagtacaaccaaccgaatctatagtgcggttccccaacattacgaacctgtgcccgttcggcgaagtgttcaacgcaacacgatttgcttctgtttacgcttggaaccggaaacgcatctccaattgcgtcgccgattacagcgttctttataattctgcatctttctccaccttcaaatgctatggtgtctctcccacaaaactcaatgacctctgttttaccaatgtgtatgcggactccttcgtcatacgcggcgacgaggtgagacaaatcgcaccagggcagactggcaagatcgctgattataattacaaactgcctgatgattttaccggatgcgttattgcttggaattctaataacctcgattccaaagttggcgggaactacaattacctctaccgattgtttcgcaaatctaaccttaagccgtttgagagagatatcagcacagagatttatcaagctggctctaccccttgcaatggagtagaaggctttaactgctattttcctcttcagtcttatggatttcaacctaccaacggggtagggtaccaaccctatagagtcgtcgtgctctcatttgaactccttcacgcccccgctaccgtgtgtgggcctaagaaatccactaatctcgttaagaataagtgtgtgaattttaatttcaatggcctgacagggaccggggttctgactgaatctaacaagaaatttctgccgttccaacaattcgggcgcgatattgcagacacgaccgacgcggtgcgcgatcctcaaacactcgaaatccttgatatcactccttgttctttcggcggtgtaagcgtcattactcctggcaccaatacctctaaccaagtggcagtactctatcaagatgtgaactgcactgaggtcccggttgcaatacatgcggatcaactcaccccaacatggcgagtgtattccacagggagcaatgtgtttcaaacgagggccggctgtctcattggggccgaacacgttaataatagttatgagtgcgatattcccattggagcgggcatttgtgccagctatcagacccaaactaactcccccgggtccgcctcatcagtcgctagccaatctattattgcgtacacaatgtccctgggagctgaaaacagcgtggcctactcaaataacagcattgcaatacccacaaattttacgatttcagtaaccactgaaatcctgcccgtctccatgaccaaaacctctgtcgactgcactatgtacatatgcggcgactccaccgagtgttccaatctccttctccaatatggaagtttctgcacgcagttgaacagggcacttacagggattgcagtcgagcaagacaagaacacccaagaagtattcgcacaagtaaaacagatctacaagacacccccaatcaaagattttggtggcttcaacttctcccaaatacttccagatccgtcaaagccatccaaacgctcattcatcgaagaccttctgttcaataaggtcacattggcggatgctggatttatcaagcaatatggggattgtttgggagatattgcagcgcgggacctgatatgcgcgcaaaagttcaatgggttgacggtgctgccccctctcctcactgacgagatgatagctcagtatacgagcgctctcctcgcgggcactatcacctcaggttggaccttcggggctggcgcggcacttcaaataccatttgctatgcaaatggcctatcgttttaatggcatcggggtgacccaaaacgtgctctatgaaaaccagaaactgatagctaatcaattcaatagtgccatcggcaaaatccaggattcattgtccagcaccgcctcagctctcgggaaattgcaagacgtcgtcaaccaaaatgctcaagcgctcaacaccctcgttaaacaactctcaagtaatttcggcgcgattagtagcgtgctgaacgatatcttgagtcgtcttgatccacctgaagcagaagtccaaatcgacaggcttattaccggacgtctgcaaagcctgcaaacctacgttacacaacaacttataagggcagccgaaataagggcttctgcaaatctggctgccacgaagatgagcgagtgtgtcctcggacaaagcaaaagagttgacttttgcggcaaagggtaccaccttatgagtttccctcagtctgcgccccatggagtagtgtttctccacgtgacttatgtaccggcacaagaaaagaactttaccacagccccagcaatatgtcacgatggaaaagcacactttccacgggaaggggttttcgtgtccaacgggacccattggtttgttactcaacgcaacttttatgaaccccaaatcataaccactgataatacatttgtctctgggaactgtgatgtcgtgatcggaatagtcaacaacacagtgtatgatccgttgcaaccagagctggattccttcaaagaagaactcgacaagtattttaagaatcacacatcaccggacgtggatcttggagacatatcaggcataaacgctagtgtggtgaatatccaaaaggagatcgacaggcttaacgaagttgcaaagaacctcaatgaatctcttatcgatttgcaagaattgggcaaatacgagcaaggctacattcctgaagcaccacgggacgggcaagcttacgtgcggaaagatggcgaatgggtgctcttgagtacctttctgggttgataa (SEQ ID NO: 119)gccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaauguuuguguuucuuguccuguugccauuggugagcucccaaugugucaaucugaccacccggacacaauugcccccugcauauacaaauucauucacgagaggaguauacuaucccgacaaaguuuuccgguccucaguccuucauuccacucaagaucuuuuccuuccauucuuuucuaacguaaccugguuccaugcaauucaugucagugggaccaacggcacgaaacgguuugauaauccaguucuuccauucaaugacggaguauauuuugcaucaacugagaaaucuaauaucauuagaggguggauuuucggaacaacucuugacuccaagacccaauccuugcucaucguuaacaaugcuacaaaugugguuauuaaggucugugaguuucaauucuguaacgaccccuuucucggcguauacuaccauaagaauaauaagucuuggauggagucugaauuucgugucuacucaucagcgaacaauuguacauuugaauaugugucccaaccauuccugauggaucucgaaggaaagcagggcaauuuuaagaaccuucgggaguucgucuuuaagaauaucgauggauacuuuaaaauauauaguaaacacacaccaaucaaucugguccgagaucucccccaggguuuuagugcucuggagccgcugguggauuugcccaucgguaucaauauuacgcgcuuccaaacauugcucgcccugcaucggaguuaccuuacgccuggcgacaguagcagcggauggaccgcuggagccgccgccuacuaugucggguaccuucaaccacgcacuuuucuccugaaauacaacgaaaaugggacaauuacagacgcuguugauugcgcacucgauccccugucagaaacaaaauguacacuuaaaucuuuuacggucgagaaagggauuuaccagacaucuaacuuccgaguacaaccaaccgaaucuauagugcgguuccccaacauuacgaaccugugcccguucggcgaaguguucaacgcaacacgauuugcuucuguuuacgcuuggaaccggaaacgcaucuccaauugcgucgccgauuacagcguucuuuauaauucugcaucuuucuccaccuucaaaugcuauggugucucucccacaaaacucaaugaccucuguuuuaccaauguguaugcggacuccuucgucauacgcggcgacgaggugagacaaaucgcaccagggcagacuggcaagaucgcugauuauaauuacaaacugccugaugauuuuaccggaugcguuauugcuuggaauucuaauaaccucgauuccaaaguuggcgggaacuacaauuaccucuaccgauuguuucgcaaaucuaaccuuaagccguuugagagagauaucagcacagagauuuaucaagcuggcucuaccccuugcaauggaguagaaggcuuuaacugcuauuuuccucuucagucuuauggauuucaaccuaccaacgggguaggguaccaacccuauagagucgucgugcucucauuugaacuccuucacgcccccgcuaccgugugugggccuaagaaauccacuaaucucguuaagaauaagugugugaauuuuaauuucaauggccugacagggaccgggguucugacugaaucuaacaagaaauuucugccguuccaacaauucgggcgcgauauugcagacacgaccgacgcggugcgcgauccucaaacacucgaaauccuugauaucacuccuuguucuuucggcgguguaagcgucauuacuccuggcaccaauaccucuaaccaaguggcaguacucuaucaagaugugaacugcacugaggucccgguugcaauacaugcggaucaacucaccccaacauggcgaguguauuccacagggagcaauguguuucaaacgagggccggcugucucauuggggccgaacacguuaauaauaguuaugagugcgauauucccauuggagcgggcauuugugccagcuaucagacccaaacuaacucccccggguccgccucaucagucgcuagccaaucuauuauugcguacacaaugucccugggagcugaaaacagcguggccuacucaaauaacagcauugcaauacccacaaauuuuacgauuucaguaaccacugaaauccugcccgucuccaugaccaaaaccucugucgacugcacuauguacauaugcggcgacuccaccgaguguuccaaucuccuucuccaauauggaaguuucugcacgcaguugaacagggcacuuacagggauugcagucgagcaagacaagaacacccaagaaguauucgcacaaguaaaacagaucuacaagacacccccaaucaaagauuuugguggcuucaacuucucccaaauacuuccagauccgucaaagccauccaaacgcucauucaucgaagaccuucuguucaauaaggucacauuggcggaugcuggauuuaucaagcaauauggggauuguuugggagauauugcagcgcgggaccugauaugcgcgcaaaaguucaauggguugacggugcugcccccucuccucacugacgagaugauagcucaguauacgagcgcucuccucgcgggcacuaucaccucagguuggaccuucggggcuggcgcggcacuucaaauaccauuugcuaugcaaauggccuaucguuuuaauggcaucggggugacccaaaacgugcucuaugaaaaccagaaacugauagcuaaucaauucaauagugccaucggcaaaauccaggauucauuguccagcaccgccucagcucucgggaaauugcaagacgucgucaaccaaaaugcucaagcgcucaacacccucguuaaacaacucucaaguaauuucggcgcgauuaguagcgugcugaacgauaucuugagucgucuugauccaccugaagcagaaguccaaaucgacaggcuuauuaccggacgucugcaaagccugcaaaccuacguuacacaacaacuuauaagggcagccgaaauaagggcuucugcaaaucuggcugccacgaagaugagcgaguguguccucggacaaagcaaaagaguugacuuuugcggcaaaggguaccaccuuaugaguuucccucagucugcgccccauggaguaguguuucuccacgugacuuauguaccggcacaagaaaagaacuuuaccacagccccagcaauaugucacgauggaaaagcacacuuuccacgggaagggguuuucguguccaacgggacccauugguuuguuacucaacgcaacuuuuaugaaccccaaaucauaaccacugauaauacauuugucucugggaacugugaugucgugaucggaauagucaacaacacaguguaugauccguugcaaccagagcuggauuccuucaaagaagaacucgacaaguauuuuaagaaucacacaucaccggacguggaucuuggagacauaucaggcauaaacgcuaguguggugaauauccaaaaggagaucgacaggcuuaacgaaguugcaaagaaccucaaugaaucucuuaucgauuugcaagaauugggcaaauacgagcaaggcuacauuccugaagcaccacgggacgggcaagcuuacgugcggaaagauggcgaaugggugcucuugaguaccuuucuggguugauaa (SEQ ID NO: 120)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICAS YQTQTNSP

A

SVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 121)WuDivS_3F_D2P_Gly_pVaxggatccgccaccatggattggacatggatattgttcttggttgcagcagctacccgggtacattccatgttcgtcttcctcgtactgctcccacttgtcagtagtcaatgtgtgaacttgactacccggacgcagttgcccccggcctacactaatagcataacgcgtggagtctattaccccgacaaggtgttcaggtcatccgtcctgtatagcactcaagatctcttcttgcccttctttagtaacgtcacttggttccatgcaatccacgtaagtggcactaatggcaccaagcgattcgacaatcccgtactcccttttaacgatggggtgtatttcgcgagcacagagaagtccaacatcatccgtggttggatcttcggcaccacactggattctaaaacccaaagcctgcttatagtaaataatgcaacaaacgtggtcattaaagtttgcgaatttcagttttgtaacgaccccttccttggcgtatattaccacaagaacaataaatcctggatggagagcgaatttagggtttacagttcagccaataattgtacattcgaatacgtaagccaacccttcttgatggacctggaaggaaagcaaggaaatttcaagaatctccgtgaattcgtgttcaagaacatagacggctattttaaaatatattcaaaacacacaccgattaacctggtacgagatcttccgcaaggattctctgcactggaaccgctggtcgatcttcctatcggcattaatatcactcggtttcaaacattgcttgctttgcatcggcgatatcttacacccggggatagctcaagtggatggactgccggggctgctgcctattacgtaggctatctccaaccacggacattcctgctgaaatataacgagaatgggacaatcacagatgctgttgactgcgctttggaccctttgagcgaaacaaagtgcacactcaaatccttcaccgtggaaaagggaatctaccaaacgtctaattttcgcgtccaaccaaccgagagcatcgtcagattcccaaacattactaatctttgcccctttggcgaagtcttcaatgctacgcgatttgcgtccgtctacgcgtggaatcggaagcgcattagcaattgcgtcgcagactattcctttctctataactctgcatccttttctacctttaaatgttatggagtcaacgggacaaagctcaatgacctttgctttacaaatgtctatgcagactcttttgtcatccgtggtgatgaggtacgacagatcgcgccaggacaaaccgggaagatcgccgattacaactacaaactgcccgacgatttcaccgggtgcgttattgcttggaactccaataatcttgatagtaaagttggcggcaactacaactacctgtatcgacttttccgtaaaagtaatctcaagccatttgaaagagacatcaacacaacaatttatcaggctggatctaccccatgcaacggcgttgaaggatttaactgctacttccctctccaaagttacggtttccaaccaacaaacggcgttggctatcaaccttatagagtcgttgtcctctcttttgagcttaaccatgccccagcgacagtgtgtgggccgaagaaaagcactaatttggttaagaataaatgtgttaactttaattttaatggattgacggggacaggggttctgacagagtctaacaagaaatttctgccgttccaacagtttgggcgagatattgcagataccacggacgccgttcgagacccccaaacacttgaaattctcgatataactccctgcagctttggcggtgtatccgttatcacgcccgggacaaataccagtaaccaagtcgcagtcctgtatcaaggcgtaaattgtacggaagtgcccgttgctatacacgctgaccaactgactcccacatggagagtctatagtactggttctaatgtgttccaaacacgagccggttgcctgatcggagccgaacatgttaacaactcatacgaatgtgacataccgattggcgccggcatttgcgccagctatcaaacgcagaccaactcaccaagaaggcgtcgcagtgtagcaagtcaatctattatagcgtataccatgtctttgggagcagaaaactccgttgcttactctaataattctattgctatcccaaccaattttacaatctcagttactaccgaaatactgccggtaagcatgactaagacatccgtggattgcactatgtacatctgtggggactcaacagagtgtagtaatttgctgcttcaatatggctccttctgcactcaactgaatcgtgctctcacgggaattgctgttgagcaagataagaatacccaggaagtgtttgcccaagtcaaacaaatttataagacaccaccaattaaagattttggtggatttaatttcagccaaatacttccagatccctcacgcagacgacggtctttcatcgaggaccttctgttcaacaaagttactctggctgatgcaggcttcattaagcagtacggtgattgtcttggagacatcgctgcgcgcgacctcatatgcgcccagaaatttaatgggctgaccgtacttccccctttgctgactgatgagatgattgcacaatacacttccgcactccttgcgggtactatcacatccgggtggacttttggagctggcgccgctcttcaaattcccttcgccatgcaaatggcgtacaggtttaatggcatcggtgtgacacagaatgtgctctatgagaaccagaaacttatcgcaaaccagttcaattcagccatcgggaaaatccaagatagtctcagtagtactgcctcagctctcggcaagctccaggatgtagtgaatcagaatgcacaagccttgaacactctcgttaaacaactttcttccaactttggtgccatcagcagtgggcctaacgatatattgagccgcttgcccaaagtggaagcggaagtccaaatagatagacttattaccggccggctgcaatctctgcaaacctatgtgactcaacaattgatccgagctgccgaaatccgtgccagtgcaaatctcgccgcgaccaagatgagcgaatgtgtcttgggacagagcaaaagagtcgatttctgcggaaaaggctaccacctgatgtctttccctcaatctgccccgcacggagtggtctttctccatgtgacttatgtgccagcccaagaaaagaactttacaaccgcaccggcaatttgccatgacggaaaggcgcatttcccccgtgagggagtctttgtgagcaacgggacccattggttcgtgacacaacgcaatttctatgagcctcagatcattaccacggacaatactttcgtgtctggcaactgtgacgtgctgataggcatcgtgaataataccgtctacgatcccttgcaacttgaactggactcattcaaagaagagctggataagtattttaagaaccatacaagccctgatgtcgatcttggggatatatcaggcataaacgcatctgttgtgaatatccaaaaggaaattgatagattgaacgaagttgccaagaacctcaatgaaagtcttatcgacctgcaagaactgggaaaatatgagcaatatataaaatggccatggagcgggcgccggagacggagaaggggtagcggcggtagtggtagcgggtacatcccagaggcacccagagatggacaagcttacgtaaggaaggacggggaatgggtgctgctcagtacatttcttggatgataa (SEQ ID NO: 122)ggauccgccaccauggauuggacauggauauuguucuugguugcagcagcuacccggguacauuccauguucgucuuccucguacugcucccacuugucaguagucaaugugugaacuugacuacccggacgcaguugcccccggccuacacuaauagcauaacgcguggagucuauuaccccgacaagguguucaggucauccguccuguauagcacucaagaucucuucuugcccuucuuuaguaacgucacuugguuccaugcaauccacguaaguggcacuaauggcaccaagcgauucgacaaucccguacucccuuuuaacgaugggguguauuucgcgagcacagagaaguccaacaucauccgugguuggaucuucggcaccacacuggauucuaaaacccaaagccugcuuauaguaaauaaugcaacaaacguggucauuaaaguuugcgaauuucaguuuuguaacgaccccuuccuuggcguauauuaccacaagaacaauaaauccuggauggagagcgaauuuaggguuuacaguucagccaauaauuguacauucgaauacguaagccaacccuucuugauggaccuggaaggaaagcaaggaaauuucaagaaucuccgugaauucguguucaagaacauagacggcuauuuuaaaauauauucaaaacacacaccgauuaaccugguacgagaucuuccgcaaggauucucugcacuggaaccgcuggucgaucuuccuaucggcauuaauaucacucgguuucaaacauugcuugcuuugcaucggcgauaucuuacacccggggauagcucaaguggauggacugccggggcugcugccuauuacguaggcuaucuccaaccacggacauuccugcugaaauauaacgagaaugggacaaucacagaugcuguugacugcgcuuuggacccuuugagcgaaacaaagugcacacucaaauccuucaccguggaaaagggaaucuaccaaacgucuaauuuucgcguccaaccaaccgagagcaucgucagauucccaaacauuacuaaucuuugccccuuuggcgaagucuucaaugcuacgcgauuugcguccgucuacgcguggaaucggaagcgcauuagcaauugcgucgcagacuauuccuuucucuauaacucugcauccuuuucuaccuuuaaauguuauggagucaacgggacaaagcucaaugaccuuugcuuuacaaaugucuaugcagacucuuuugucauccguggugaugagguacgacagaucgcgccaggacaaaccgggaagaucgccgauuacaacuacaaacugcccgacgauuucaccgggugcguuauugcuuggaacuccaauaaucuugauaguaaaguuggcggcaacuacaacuaccuguaucgacuuuuccguaaaaguaaucucaagccauuugaaagagacaucaacacaacaauuuaucaggcuggaucuaccccaugcaacggcguugaaggauuuaacugcuacuucccucuccaaaguuacgguuuccaaccaacaaacggcguuggcuaucaaccuuauagagucguuguccucucuuuugagcuuaaccaugccccagcgacagugugugggccgaagaaaagcacuaauuugguuaagaauaaauguguuaacuuuaauuuuaauggauugacggggacagggguucugacagagucuaacaagaaauuucugccguuccaacaguuugggcgagauauugcagauaccacggacgccguucgagacccccaaacacuugaaauucucgauauaacucccugcagcuuuggcgguguauccguuaucacgcccgggacaaauaccaguaaccaagucgcaguccuguaucaaggcguaaauuguacggaagugcccguugcuauacacgcugaccaacugacucccacauggagagucuauaguacugguucuaauguguuccaaacacgagccgguugccugaucggagccgaacauguuaacaacucauacgaaugugacauaccgauuggcgccggcauuugcgccagcuaucaaacgcagaccaacucaccaagaaggcgucgcaguguagcaagucaaucuauuauagcguauaccaugucuuugggagcagaaaacuccguugcuuacucuaauaauucuauugcuaucccaaccaauuuuacaaucucaguuacuaccgaaauacugccgguaagcaugacuaagacauccguggauugcacuauguacaucuguggggacucaacagaguguaguaauuugcugcuucaauauggcuccuucugcacucaacugaaucgugcucucacgggaauugcuguugagcaagauaagaauacccaggaaguguuugcccaagucaaacaaauuuauaagacaccaccaauuaaagauuuugguggauuuaauuucagccaaauacuuccagaucccucacgcagacgacggucuuucaucgaggaccuucuguucaacaaaguuacucuggcugaugcaggcuucauuaagcaguacggugauugucuuggagacaucgcugcgcgcgaccucauaugcgcccagaaauuuaaugggcugaccguacuucccccuuugcugacugaugagaugauugcacaauacacuuccgcacuccuugcggguacuaucacauccggguggacuuuuggagcuggcgccgcucuucaaauucccuucgccaugcaaauggcguacagguuuaauggcaucggugugacacagaaugugcucuaugagaaccagaaacuuaucgcaaaccaguucaauucagccaucgggaaaauccaagauagucucaguaguacugccucagcucucggcaagcuccaggauguagugaaucagaaugcacaagccuugaacacucucguuaaacaacuuucuuccaacuuuggugccaucagcagugggccuaacgauauauugagccgcuugcccaaaguggaagcggaaguccaaauagauagacuuauuaccggccggcugcaaucucugcaaaccuaugugacucaacaauugauccgagcugccgaaauccgugccagugcaaaucucgccgcgaccaagaugagcgaaugugucuugggacagagcaaaagagucgauuucugcggaaaaggcuaccaccugaugucuuucccucaaucugccccgcacggaguggucuuucuccaugugacuuaugugccagcccaagaaaagaacuuuacaaccgcaccggcaauuugccaugacggaaaggcgcauuucccccgugagggagucuuugugagcaacgggacccauugguucgugacacaacgcaauuucuaugagccucagaucauuaccacggacaauacuuucgugucuggcaacugugacgugcugauaggcaucgugaauaauaccgucuacgaucccuugcaacuugaacuggacucauucaaagaagagcuggauaaguauuuuaagaaccauacaagcccugaugucgaucuuggggauauaucaggcauaaacgcaucuguugugaauauccaaaaggaaauugauagauugaacgaaguugccaagaaccucaaugaaagucuuaucgaccugcaagaacugggaaaauaugagcaauauauaaaauggccauggagcgggcgccggagacggagaagggguagcggcgguagugguagcggguacaucccagaggcacccagagauggacaagcuuacguaaggaaggacggggaaugggugcugcucaguacauuucuuggaugauaa (SEQ ID NO: 123)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGV

T KLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDI

T

IYQAGSTPCNGVEGFNCYFPLQSYGFQPT NGVGYQPYRVVVLSFEL

HAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPS

RSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVK QLSSNFGAISS

NDILSRL

KVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 124) WuS_3F_2P_NoTri_pVaxggatccgccaccatggactggacgtggattttgtttcttgtcgctgcagctacccgggttcactccatgtttgttttcctggtgctccttccccttgtaagctcacaatgcgtcaatttgaccacgcgtacacaactgcccccagcatatactaattctttcacacgcggggtctactatcccgataaagtctttagaagtagcgtacttcatagtacccaagatttgtttctgcccttcttcagcaatgtcacgtggtttcatgcgatccatgtatccggcacgaacgggacaaaacgatttgataaccccgtgctccccttcaacgacggggtttatttcgccagcaccgagaaatcaaatattatcaggggctggattttcgggacaacacttgattccaagacacaatctcttcttatcgtgaataatgcaactaatgtggtgatcaaggtttgcgagttccaattttgtaatgacccttttcttggcgtgtactatcataagaataacaagagttggatggaatcagagttccgggtctacagcagtgctaacaattgtacgtttgaatacgtttctcagccttttctgatggaccttgaaggtaagcaaggcaatttcaagaacttgcgggaatttgtcttcaagaacatagatggctattttaagatatatagcaaacacactcccataaatctcgtcagagatcttccacagggctttagcgccctggaaccattggttgatttgccaattggaataaacataactcgattccaaaccttgctcgcactccatcggagctacctgacgcctggagattcctcttccggctggactgccggagcagcagcttattatgtaggctacttgcaaccccgcacgttcctgctcaaatataatgaaaatggcactataaccgatgcggtagactgcgctcttgatcccctgagtgaaactaaatgtacgttgaaaagctttactgtagagaaaggcatctatcagactagtaactttagggtgcaacccacggagtccattgtacggttcccaaacattaccaacctctgtccattcggagaagtgtttaatgccacaagattcgcttcagtgtatgcctggaaccggaaacgcatctcaaattgcgttgccgattattcagtactttacaactcagccagtttctctacttttaagtgctatggcgtttccccgacgaagctcaatgatctgtgctttactaacgtttacgcagactctttcgtcatcagaggcgatgaagtcaggcaaatagctcctggtcaaaccggcaagatcgccgactacaactataaactgcccgatgatttcactgggtgtgtgatcgcgtggaattccaataatttggactctaaggtaggtggcaactataactacctctatcgactcttccgaaaatccaaccttaagccgtttgaacgcgatattagtaccgaaatataccaagccgggtctacaccctgtaacggcgttgaaggtttcaattgttactttccactgcagagttatggctttcaacccaccaacggggttggctatcagccctatagggttgtggtcctcagttttgagcttctgcatgcaccagcaaccgtgtgcggacctaagaagtcaacaaatctcgtgaagaacaagtgtgttaatttcaatttcaatggccttacagggaccggagtgcttacagaaagcaataagaagttcttgccctttcaacagttcggcagggacatagcggacacgacagatgcagttcgagacccgcaaactctcgaaattctggatatcacaccttgcagttttggtggcgtgtctgttatcacaccaggcaccaacacttccaaccaggtggcagttttgtaccaggatgttaattgtacagaggtcccagtggcaatacacgctgaccaactgactccaacttggagagtctactctacaggctcaaacgtcttccaaacacgggcggggtgtctgatcggagcagaacacgttaataacagttacgagtgtgatatcccgataggagctggtatttgcgcttcataccagacgcaaacgaactcaccacgaagacgccggtcagttgcatcacaatccattattgcatacaccatgtcactcggagcggagaattctgtagcatacagtaacaatagtatcgcaatacctacgaactttaccatttccgtcacaactgaaatcttgcccgtctcaatgacaaagacaagcgtagattgtacaatgtatatttgcggagattcaacagagtgctccaacctgctgctccagtacggtagtttctgtacccagctcaatagggccctcaccggaattgcagttgaacaagacaagaacacccaagaagtgtttgcacaagtcaaacaaatctataaaacacccccaatcaaagatttcggtggcttcaacttttcacaaattctccctgatcctagccgccgccgcagatcattcatcgaagacttgctcttcaataaggttaccctggcagacgccggttttattaaacaatacggagattgcctcggtgacatcgccgctagagaccttatctgtgcccaaaagttcaacggactcaccgtgctgcccccattgctgaccgatgaaatgattgctcaatatacatctgcgctcctcgcagggaccattacttcagggtggacttttggggctggcgccgcattgcagattcccttcgccatgcagatggcatataggtttaacggcattggagttacccaaaatgtactctacgagaaccaaaagctgattgcaaatcagttcaacagtgcaataggcaaaatacaagactctctgtcttcaaccgccagcgctcttggaaagctccaagatgttgttaatcaaaatgcccaagcgttgaataccctcgtgaagcaactctccagcaattttggtgccatctctagcgtgctgaacgacattctgtcacggctcgatcccccggaagccgaggtacaaattgaccgattgataaccgggcgactccaaagccttcagacctacgttacacaacagctcattcgcgctgcagaaattagagcctctgcaaatcttgcagctacaaagatgtcagagtgcgttctcggtcaaagcaaaagagtggatttctgcggaaaggggtaccacctcatgagtttcccacagagtgcccctcatggcgtagtctttcttcatgttacttatgtaccagcccaagaaaagaatttcactacagcacccgcgatttgtcatgatggcaaagcgcacttccctcgggaaggcgtgttcgtgtctaatggaacacattggttcgtgacgcaacggaatttctacgagccccaaattatcactactgataacaccttcgtctccggaaactgcgatgttgttattggcattgtcaacaataccgtttacgacccgctccaacctgagctggattcatttaaagaggaattggacaaatattttaagaatcatacctctccagacgtggatttgggtgacattagcggaataaatgcatctgtggtcaatatccaaaaggaaattgataggctgaacgaggtcgccaagaatttgaacgaatctttgattgatcttcaagaacttggcaagtatgaacaatacataaaatggccctggtgatag (SEQ IDNO: 125)ggauccgccaccauggacuggacguggauuuuguuucuugucgcugcagcuacccggguucacuccauguuuguuuuccuggugcuccuuccccuuguaagcucacaaugcgucaauuugaccacgcguacacaacugcccccagcauauacuaauucuuucacacgcggggucuacuaucccgauaaagucuuuagaaguagcguacuucauaguacccaagauuuguuucugcccuucuucagcaaugucacgugguuucaugcgauccauguauccggcacgaacgggacaaaacgauuugauaaccccgugcuccccuucaacgacgggguuuauuucgccagcaccgagaaaucaaauauuaucaggggcuggauuuucgggacaacacuugauuccaagacacaaucucuucuuaucgugaauaaugcaacuaauguggugaucaagguuugcgaguuccaauuuuguaaugacccuuuucuuggcguguacuaucauaagaauaacaagaguuggauggaaucagaguuccgggucuacagcagugcuaacaauuguacguuugaauacguuucucagccuuuucugauggaccuugaagguaagcaaggcaauuucaagaacuugcgggaauuugucuucaagaacauagauggcuauuuuaagauauauagcaaacacacucccauaaaucucgucagagaucuuccacagggcuuuagcgcccuggaaccauugguugauuugccaauuggaauaaacauaacucgauuccaaaccuugcucgcacuccaucggagcuaccugacgccuggagauuccucuuccggcuggacugccggagcagcagcuuauuauguaggcuacuugcaaccccgcacguuccugcucaaauauaaugaaaauggcacuauaaccgaugcgguagacugcgcucuugauccccugagugaaacuaaauguacguugaaaagcuuuacuguagagaaaggcaucuaucagacuaguaacuuuagggugcaacccacggaguccauuguacgguucccaaacauuaccaaccucuguccauucggagaaguguuuaaugccacaagauucgcuucaguguaugccuggaaccggaaacgcaucucaaauugcguugccgauuauucaguacuuuacaacucagccaguuucucuacuuuuaagugcuauggcguuuccccgacgaagcucaaugaucugugcuuuacuaacguuuacgcagacucuuucgucaucagaggcgaugaagucaggcaaauagcuccuggucaaaccggcaagaucgccgacuacaacuauaaacugcccgaugauuucacugggugugugaucgcguggaauuccaauaauuuggacucuaagguagguggcaacuauaacuaccucuaucgacucuuccgaaaauccaaccuuaagccguuugaacgcgauauuaguaccgaaauauaccaagccgggucuacacccuguaacggcguugaagguuucaauuguuacuuuccacugcagaguuauggcuuucaacccaccaacgggguuggcuaucagcccuauaggguugugguccucaguuuugagcuucugcaugcaccagcaaccgugugcggaccuaagaagucaacaaaucucgugaagaacaaguguguuaauuucaauuucaauggccuuacagggaccggagugcuuacagaaagcaauaagaaguucuugcccuuucaacaguucggcagggacauagcggacacgacagaugcaguucgagacccgcaaacucucgaaauucuggauaucacaccuugcaguuuugguggcgugucuguuaucacaccaggcaccaacacuuccaaccagguggcaguuuuguaccaggauguuaauuguacagaggucccaguggcaauacacgcugaccaacugacuccaacuuggagagucuacucuacaggcucaaacgucuuccaaacacgggcggggugucugaucggagcagaacacguuaauaacaguuacgagugugauaucccgauaggagcugguauuugcgcuucauaccagacgcaaacgaacucaccacgaagacgccggucaguugcaucacaauccauuauugcauacaccaugucacucggagcggagaauucuguagcauacaguaacaauaguaucgcaauaccuacgaacuuuaccauuuccgucacaacugaaaucuugcccgucucaaugacaaagacaagcguagauuguacaauguauauuugcggagauucaacagagugcuccaaccugcugcuccaguacgguaguuucuguacccagcucaauagggcccucaccggaauugcaguugaacaagacaagaacacccaagaaguguuugcacaagucaaacaaaucuauaaaacacccccaaucaaagauuucgguggcuucaacuuuucacaaauucucccugauccuagccgccgccgcagaucauucaucgaagacuugcucuucaauaagguuacccuggcagacgccgguuuuauuaaacaauacggagauugccucggugacaucgccgcuagagaccuuaucugugcccaaaaguucaacggacucaccgugcugcccccauugcugaccgaugaaaugauugcucaauauacaucugcgcuccucgcagggaccauuacuucaggguggacuuuuggggcuggcgccgcauugcagauucccuucgccaugcagauggcauauagguuuaacggcauuggaguuacccaaaauguacucuacgagaaccaaaagcugauugcaaaucaguucaacagugcaauaggcaaaauacaagacucucugucuucaaccgccagcgcucuuggaaagcuccaagauguuguuaaucaaaaugcccaagcguugaauacccucgugaagcaacucuccagcaauuuuggugccaucucuagcgugcugaacgacauucugucacggcucgaucccccggaagccgagguacaaauugaccgauugauaaccgggcgacuccaaagccuucagaccuacguuacacaacagcucauucgcgcugcagaaauuagagccucugcaaaucuugcagcuacaaagaugucagagugcguucucggucaaagcaaaagaguggauuucugcggaaagggguaccaccucaugaguuucccacagagugccccucauggcguagucuuucuucauguuacuuauguaccagcccaagaaaagaauuucacuacagcacccgcgauuugucaugauggcaaagcgcacuucccucgggaaggcguguucgugucuaauggaacacauugguucgugacgcaacggaauuucuacgagccccaaauuaucacuacugauaacaccuucgucuccggaaacugcgauguuguuauuggcauugucaacaauaccguuuacgacccgcuccaaccugagcuggauucauuuaaagaggaauuggacaaauauuuuaagaaucauaccucuccagacguggauuugggugacauuagcggaauaaaugcaucuguggucaauauccaaaaggaaauugauaggcugaacgaggucgccaagaauuugaacgaaucuuugauugaucuucaagaacuuggcaaguaugaacaauacauaaaauggcccuggugauag (SEQ ID NO: 126)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICAS YQTQTNSPRR

RSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPS

RSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLD

EAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPW** (SEQ ID NO: 127) WuS_3F_D2P_Gly_pVaxggatccgccaccatggattggacatggatattgttcttggttgcagcagctacccgggtacattccatgttcgtcttcctcgtactgctcccacttgtcagtagtcaatgtgtgaacttgactacccggacgcagttgcccccggcctacactaatagctttacgcgtggagtctattaccccgacaaggtgttcaggtcatccgtcctgcatagcactcaagatctcttcttgcccttctttagtaacgtcacttggttccatgcaatccacgtaagtggcactaatggcaccaagcgattcgacaatcccgtactcccttttaacgatggggtgtatttcgcgagcacagagaagtccaacatcatccgtggttggatcttcggcaccacactggattctaaaacccaaagcctgcttatagtaaataatgcaacaaacgtggtcattaaagtttgcgaatttcagttttgtaacgaccccttccttggcgtatattaccacaagaacaataaatcctggatggagagcgaatttagggtttacagttcagccaataattgtacattcgaatacgtaagccaacccttcttgatggacctggaaggaaagcaaggaaatttcaagaatctccgtgaattcgtgttcaagaacatagacggctattttaaaatatattcaaaacacacaccgattaacctggtacgagatcttccgcaaggattctctgcactggaaccgctggtcgatcttcctatcggcattaatatcactcggtttcaaacattgcttgctttgcatcggagttatcttacacccggggatagctcaagtggatggactgccggggctgctgcctattacgtaggctatctccaaccacggacattcctgctgaaatataacgagaatgggacaatcacagatgctgttgactgcgctttggaccctttgagcgaaacaaagtgcacactcaaatccttcaccgtggaaaagggaatctaccaaacgtctaattttcgcgtccaaccaaccgagagcatcgtcagattcccaaacattactaatctttgcccctttggcgaagtcttcaatgctacgcgatttgcgtccgtctacgcgtggaatcggaagcgcattagcaattgcgtcgcagactattccgtgctctataactctgcatccttttctacctttaaatgttatggagtcaacgggacaaagctcaatgacctttgctttacaaatgtctatgcagactcttttgtcatccgtggtgatgaggtacgacagatcgcgccaggacaaaccgggaagatcgccgattacaactacaaactgcccgacgatttcaccgggtgcgttattgcttggaactccaataatcttgatagtaaagttggcggcaactacaactacctgtatcgacttttccgtaaaagtaatctcaagccatttgaaagagacatcaacacaacaatttatcaggctggatctaccccatgcaacggcgttgaaggatttaactgctacttccctctccaaagttacggtttccaaccaacaaacggcgttggctatcaaccttatagagtcgttgtcctctcttttgagcttaaccatgccccagcgacagtgtgtgggccgaagaaaagcactaatttggttaagaataaatgtgttaactttaattttaatggattgacggggacaggggttctgacagagtctaacaagaaatttctgccgttccaacagtttgggcgagatattgcagataccacggacgccgttcgagacccccaaacacttgaaattctcgatataactccctgcagctttggcggtgtatccgttatcacgcccgggacaaataccagtaaccaagtcgcagtcctgtatcaagacgtaaattgtacggaagtgcccgttgctatacacgctgaccaactgactcccacatggagagtctatagtactggttctaatgtgttccaaacacgagccggttgcctgatcggagccgaacatgttaacaactcatacgaatgtgacataccgattggcgccggcatttgcgccagctatcaaacgcagaccaactcaccaagaaggcgtcgcagtgtagcaagtcaatctattatagcgtataccatgtctttgggagcagaaaactccgttgcttactctaataattctattgctatcccaaccaattttacaatctcagttactaccgaaatactgccggtaagcatgactaagacatccgtggattgcactatgtacatctgtggggactcaacagagtgtagtaatttgctgcttcaatatggctccttctgcactcaactgaatcgtgctctcacgggaattgctgttgagcaagataagaatacccaggaagtgtttgcccaagtcaaacaaatttataagacaccaccaattaaagattttggtggatttaatttcagccaaatacttccagatccctcacgcagacgacggtctttcatcgaggaccttctgttcaacaaagttactctggctgatgcaggcttcattaagcagtacggtgattgtcttggagacatcgctgcgcgcgacctcatatgcgcccagaaatttaatgggctgaccgtacttccccctttgctgactgatgagatgattgcacaatacacttccgcactccttgcgggtactatcacatccgggtggacttttggagctggcgccgctcttcaaattcccttcgccatgcaaatggcgtacaggtttaatggcatcggtgtgacacagaatgtgctctatgagaaccagaaacttatcgcaaaccagttcaattcagccatcgggaaaatccaagatagtctcagtagtactgcctcagctctcggcaagctccaggatgtagtgaatcagaatgcacaagccttgaacactctcgttaaacaactttcttccaactttggtgccatcagcagtgggcctaacgatatattgagccgcttgcccaaagtggaagcggaagtccaaatagatagacttattaccggccggctgcaatctctgcaaacctatgtgactcaacaattgatccgagctgccgaaatccgtgccagtgcaaatctcgccgcgaccaagatgagcgaatgtgtcttgggacagagcaaaagagtcgatttctgcggaaaaggctaccacctgatgtctttccctcaatctgccccgcacggagtggtctttctccatgtgacttatgtgccagcccaagaaaagaactttacaaccgcaccggcaatttgccatgacggaaaggcgcatttcccccgtgagggagtctttgtgagcaacgggacccattggttcgtgacacaacgcaatttctatgagcctcagatcattaccacggacaatactttcgtgtctggcaactgtgacgtggtcataggcatcgtgaataataccgtctacgatcccttgcaacccgaactggactcattcaaagaagagctggataagtattttaagaaccatacaagccctgatgtcgatcttggggatatatcaggcataaacgcatctgttgtgaatatccaaaaggaaattgatagattgaacgaagttgccaagaacctcaatgaaagtcttatcgacctgcaagaactgggaaaatatgagcaatatataaaatggccatggagcgggcgccggagacggagaaggggtagcggcggtagtggtagcgggtacatcccagaggcacccagagatggacaagcttacgtaaggaaggacggggaatgggtgctgctcagtacatttcttggatgataa (SEQ ID NO: 128)ggauccgccaccauggauuggacauggauauuguucuugguugcagcagcuacccggguacauuccauguucgucuuccucguacugcucccacuugucaguagucaaugugugaacuugacuacccggacgcaguugcccccggccuacacuaauagcuuuacgcguggagucuauuaccccgacaagguguucaggucauccguccugcauagcacucaagaucucuucuugcccuucuuuaguaacgucacuugguuccaugcaauccacguaaguggcacuaauggcaccaagcgauucgacaaucccguacucccuuuuaacgaugggguguauuucgcgagcacagagaaguccaacaucauccgugguuggaucuucggcaccacacuggauucuaaaacccaaagccugcuuauaguaaauaaugcaacaaacguggucauuaaaguuugcgaauuucaguuuuguaacgaccccuuccuuggcguauauuaccacaagaacaauaaauccuggauggagagcgaauuuaggguuuacaguucagccaauaauuguacauucgaauacguaagccaacccuucuugauggaccuggaaggaaagcaaggaaauuucaagaaucuccgugaauucguguucaagaacauagacggcuauuuuaaaauauauucaaaacacacaccgauuaaccugguacgagaucuuccgcaaggauucucugcacuggaaccgcuggucgaucuuccuaucggcauuaauaucacucgguuucaaacauugcuugcuuugcaucggaguuaucuuacacccggggauagcucaaguggauggacugccggggcugcugccuauuacguaggcuaucuccaaccacggacauuccugcugaaauauaacgagaaugggacaaucacagaugcuguugacugcgcuuuggacccuuugagcgaaacaaagugcacacucaaauccuucaccguggaaaagggaaucuaccaaacgucuaauuuucgcguccaaccaaccgagagcaucgucagauucccaaacauuacuaaucuuugccccuuuggcgaagucuucaaugcuacgcgauuugcguccgucuacgcguggaaucggaagcgcauuagcaauugcgucgcagacuauuccgugcucuauaacucugcauccuuuucuaccuuuaaauguuauggagucaacgggacaaagcucaaugaccuuugcuuuacaaaugucuaugcagacucuuuugucauccguggugaugagguacgacagaucgcgccaggacaaaccgggaagaucgccgauuacaacuacaaacugcccgacgauuucaccgggugcguuauugcuuggaacuccaauaaucuugauaguaaaguuggcggcaacuacaacuaccuguaucgacuuuuccguaaaaguaaucucaagccauuugaaagagacaucaacacaacaauuuaucaggcuggaucuaccccaugcaacggcguugaaggauuuaacugcuacuucccucuccaaaguuacgguuuccaaccaacaaacggcguuggcuaucaaccuuauagagucguuguccucucuuuugagcuuaaccaugccccagcgacagugugugggccgaagaaaagcacuaauuugguuaagaauaaauguguuaacuuuaauuuuaauggauugacggggacagggguucugacagagucuaacaagaaauuucugccguuccaacaguuugggcgagauauugcagauaccacggacgccguucgagacccccaaacacuugaaauucucgauauaacucccugcagcuuuggcgguguauccguuaucacgcccgggacaaauaccaguaaccaagucgcaguccuguaucaagacguaaauuguacggaagugcccguugcuauacacgcugaccaacugacucccacauggagagucuauaguacugguucuaauguguuccaaacacgagccgguugccugaucggagccgaacauguuaacaacucauacgaaugugacauaccgauuggcgccggcauuugcgccagcuaucaaacgcagaccaacucaccaagaaggcgucgcaguguagcaagucaaucuauuauagcguauaccaugucuuugggagcagaaaacuccguugcuuacucuaauaauucuauugcuaucccaaccaauuuuacaaucucaguuacuaccgaaauacugccgguaagcaugacuaagacauccguggauugcacuauguacaucuguggggacucaacagaguguaguaauuugcugcuucaauauggcuccuucugcacucaacugaaucgugcucucacgggaauugcuguugagcaagauaagaauacccaggaaguguuugcccaagucaaacaaauuuauaagacaccaccaauuaaagauuuugguggauuuaauuucagccaaauacuuccagaucccucacgcagacgacggucuuucaucgaggaccuucuguucaacaaaguuacucuggcugaugcaggcuucauuaagcaguacggugauugucuuggagacaucgcugcgcgcgaccucauaugcgcccagaaauuuaaugggcugaccguacuucccccuuugcugacugaugagaugauugcacaauacacuuccgcacuccuugcggguacuaucacauccggguggacuuuuggagcuggcgccgcucuucaaauucccuucgccaugcaaauggcguacagguuuaauggcaucggugugacacagaaugugcucuaugagaaccagaaacuuaucgcaaaccaguucaauucagccaucgggaaaauccaagauagucucaguaguacugccucagcucucggcaagcuccaggauguagugaaucagaaugcacaagccuugaacacucucguuaaacaacuuucuuccaacuuuggugccaucagcagugggccuaacgauauauugagccgcuugcccaaaguggaagcggaaguccaaauagauagacuuauuaccggccggcugcaaucucugcaaaccuaugugacucaacaauugauccgagcugccgaaauccgugccagugcaaaucucgccgcgaccaagaugagcgaaugugucuugggacagagcaaaagagucgauuucugcggaaaaggcuaccaccugaugucuuucccucaaucugccccgcacggaguggucuuucuccaugugacuuaugugccagcccaagaaaagaacuuuacaaccgcaccggcaauuugccaugacggaaaggcgcauuucccccgugagggagucuuugugagcaacgggacccauugguucgugacacaacgcaauuucuaugagccucagaucauuaccacggacaauacuuucgugucuggcaacugugacguggucauaggcaucgugaauaauaccgucuacgaucccuugcaacccgaacuggacucauucaaagaagagcuggauaaguauuuuaagaaccauacaagcccugaugucgaucuuggggauauaucaggcauaaacgcaucuguugugaauauccaaaaggaaauugauagauugaacgaaguugccaagaaccucaaugaaagucuuaucgaccugcaagaacugggaaaauaugagcaauauauaaaauggccauggagcgggcgccggagacggagaagggguagcggcgguagugguagcggguacaucccagaggcacccagagauggacaagcuuacguaaggaaggacggggaaugggugcugcucaguacauuucuuggaugauaa (SEQ ID NO: 129)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGV

T KLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDI

T

IYQAGSTPCNGVEGFNCYFPLQSYGFQPT NGVGYQPYRVVVLSFEL

HAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICA SYQTQTNSPRR

RSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPS

RSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVK QLSSNFGAISS

NDILSRL

KVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 130) WuS_3F_D2P_rB_pVaxggatccgccaccatggattggacatggattctgtttctcgttgccgccgctacgcgcgtgcacagcatgttcgttttcctcgtactgttgcctctggtatcatctcaatgtgtgaatcttacgacgaggacgcaactgccaccagcttacaccaactcttttactagaggggtctattaccccgacaaggttttcagatcctcagtgctgcatagtacacaagatttgtttcttcccttcttctccaatgtcacttggtttcacgctatccacgtgtccggcactaatggaacgaagcggtttgataacccggtacttccatttaatgacggggtatactttgcaagcaccgagaaaagtaatatcattcgtgggtggatctttggcactacactcgactccaagacacaatcccttttgatagtaaataatgctacaaatgtggttataaaggtgtgcgagtttcaattctgcaatgaccccttcctcggtgtctattatcacaagaacaataaatcttggatggaaagtgagttcagggtatacagctctgcaaacaactgcacatttgagtatgtgagccaaccgtttcttatggatctggaaggtaagcagggtaactttaagaaccttcgggagttcgtctttaagaatatagatggctattttaaaatctatagtaaacacactcctattaaccttgtgcgcgatctccctcaagggttctctgcattggaaccgcttgttgatttgcctataggaatcaatattacacgatttcaaacactcctcgctctccataggagctaccttaccccaggcgactcaagctctggttggacggcaggagctgcagcatactatgtgggttatcttcagccgcggacattcttgcttaagtataatgagaatggaactatcactgacgctgttgactgtgccctggaccctctttcagaaacaaaatgtactctcaaatcattcaccgtggagaagggaatatatcaaacaagtaactttagggtccaacctaccgagagcatcgtgcgattccctaatattaccaatttgtgtcccttcggtgaagtcttcaacgcgacccgctttgctagcgtctatgcgtggaacaggaagcgaattagcaactgtgttgcagattacagtgtgctgtacaatagtgcgagcttttccacgttcaaatgctatggcgtgaaacctaccaaacttaatgatttgtgcttcactaatgtttatgctgacagcttcgttatccggggtgatgaggtgaggcagattgcccccggtcaaactggcaaaattgccgactacaattacaagctccctgatgattttactggatgtgtcatagcgtggaattccaataatctggactctaaggttggtggtaattataactatctctaccgcctgtttcgtaagagcaatctgaaaccctttgaaagagatatttggactgagatatatcaagctggctcaactccttgcaacggggtcgaaggtttcaattgttactttccacttcaatcatacgggtttcaaccaactaacggtgtaggttatcaaccctatcgggtggttgtcctgagctttgagctgaaccatgccccggctacagtatgcggcccaaagaaatccactaacttggtcaagaacaaatgcgtcaactttaactttaacggactcacggggacaggagtccttaccgaatccaacaagaaattcttgcctttccaacaatttggacgagacattgcggataccacagacgcagtacgcgacccacagactcttgaaatcctcgacataacaccctgcagtttcggcggtgtaagtgtcattaccccaggcactaatacgagcaaccaagtggcggtgctctaccaagacgttaattgcactgaggtcccagtggctattcacgctgaccaacttacacccacatggagagtgtatagtacaggctcaaacgtcttccagacacgggcggggtgccttattggagcagaacatgttaacaattcctatgaatgcgatatcccgattggagccgggatctgtgctagctatcaaacccaaacaaatagccccagacgtcgacgttccgtggctagtcaaagcatcatcgcctacactatgagtcttggggccgaaaattccgttgcttacagtaacaacagtatcgctatccccaccaattttactattagtgtaactacagagattctgccggtttccatgacaaagacttccgtggattgtacgatgtatatttgcggcgacagcacagagtgcagcaatctgctgctgcaatacggtagtttctgcacccaattgaaccgtgctctgacgggaattgcagttgagcaggacaagaatactcaagaagtatttgcacaagtcaaacagatatacaagacgcccccgattaaagatttcggcgggtttaactttagccaaattcttccggaccccagcagacgccgccgaagctttattgaggacctgctgtttaataaagttacccttgctgatgctggttttatcaagcaatacggagattgcctgggagatatcgccgccagggatttgatctgtgcgcaaaagtttaacggccttaccgttctcccgccccttctgaccgatgaaatgatagcccaatacacttccgcactcctggcaggcacaattacttccggctggacgtttggggccggggcagccttgcaaattccgtttgctatgcaaatggcatatcgtttcaatggtatcggcgtaacacaaaatgtcctttatgagaaccagaaactcattgctaatcagtttaattccgctatcggcaagattcaagacagtctcagcagcacggcgagcgcacttggtaaacttcaagacgttgtcaaccagaatgctcaagccctgaacactctggtaaaacaacttagctctaatttcggtgcaattagctccggtccgaacgatattctgtcacggctcccgaaagtcgaagccgaagtccagatcgataggctgatcacagggcgcttgcagagtctccaaacctacgtgacgcaacaactcattcgggcggctgaaattcgtgcaagcgctaatctggccgctaccaaaatgagtgagtgtgttctcggtcaatcaaagagggttgacttttgcggcaaaggatatcatttgatgagttttccgcaatctgcccctcatggggtagtatttctgcacgtaacttatgtaccagcacaagaaaagaacttcaccacggccccagcaatatgccacgatggcaaagctcatttccctcgcgaaggggtctttgtaagcaatggaacccactggtttgtcacacaacgcaacttttatgagcctcaaatcattacaaccgataacacttttgtctccgggaactgcgacgtggtgattggaatcgtcaacaacactgtctatgatcccctgcaacctgaactggattcctttaaagaagagcttgataagtatttcaagaaccataccagccccgacgtcgatttgggagatattagtgggattaatgctagcgttgttaatatacaaaaggaaatagatcgattgaatgaagtggccaagaatctgaatgagtctctgattgacctgcaggagctcggaaagtatgagcaatatataaaatggccctggtcaggccgcaggcgtcggcggcgcggtagcggcggttcaggatctgggtatatacctgaggccccacgagatgggcaggcttatgtacggaaagatggagaatgggtgttgctgagtactttcctcgggtaataa (SEQ ID NO: 131)ggauccgccaccauggauuggacauggauucuguuucucguugccgccgcuacgcgcgugcacagcauguucguuuuccucguacuguugccucugguaucaucucaaugugugaaucuuacgacgaggacgcaacugccaccagcuuacaccaacucuuuuacuagaggggucuauuaccccgacaagguuuucagauccucagugcugcauaguacacaagauuuguuucuucccuucuucuccaaugucacuugguuucacgcuauccacguguccggcacuaauggaacgaagcgguuugauaacccgguacuuccauuuaaugacgggguauacuuugcaagcaccgagaaaaguaauaucauucguggguggaucuuuggcacuacacucgacuccaagacacaaucccuuuugauaguaaauaaugcuacaaaugugguuauaaaggugugcgaguuucaauucugcaaugaccccuuccucggugucuauuaucacaagaacaauaaaucuuggauggaaagugaguucaggguauacagcucugcaaacaacugcacauuugaguaugugagccaaccguuucuuauggaucuggaagguaagcaggguaacuuuaagaaccuucgggaguucgucuuuaagaauauagauggcuauuuuaaaaucuauaguaaacacacuccuauuaaccuugugcgcgaucucccucaaggguucucugcauuggaaccgcuuguugauuugccuauaggaaucaauauuacacgauuucaaacacuccucgcucuccauaggagcuaccuuaccccaggcgacucaagcucugguuggacggcaggagcugcagcauacuauguggguuaucuucagccgcggacauucuugcuuaaguauaaugagaauggaacuaucacugacgcuguugacugugcccuggacccucuuucagaaacaaaauguacucucaaaucauucaccguggagaagggaauauaucaaacaaguaacuuuaggguccaaccuaccgagagcaucgugcgauucccuaauauuaccaauuugugucccuucggugaagucuucaacgcgacccgcuuugcuagcgucuaugcguggaacaggaagcgaauuagcaacuguguugcagauuacagugugcuguacaauagugcgagcuuuuccacguucaaaugcuauggcgugaaaccuaccaaacuuaaugauuugugcuucacuaauguuuaugcugacagcuucguuauccggggugaugaggugaggcagauugcccccggucaaacuggcaaaauugccgacuacaauuacaagcucccugaugauuuuacuggaugugucauagcguggaauuccaauaaucuggacucuaagguuggugguaauuauaacuaucucuaccgccuguuucguaagagcaaucugaaacccuuugaaagagauauuuggacugagauauaucaagcuggcucaacuccuugcaacggggucgaagguuucaauuguuacuuuccacuucaaucauacggguuucaaccaacuaacgguguagguuaucaacccuaucgggugguuguccugagcuuugagcugaaccaugccccggcuacaguaugcggcccaaagaaauccacuaacuuggucaagaacaaaugcgucaacuuuaacuuuaacggacucacggggacaggaguccuuaccgaauccaacaagaaauucuugccuuuccaacaauuuggacgagacauugcggauaccacagacgcaguacgcgacccacagacucuugaaauccucgacauaacacccugcaguuucggcgguguaagugucauuaccccaggcacuaauacgagcaaccaaguggcggugcucuaccaagacguuaauugcacugaggucccaguggcuauucacgcugaccaacuuacacccacauggagaguguauaguacaggcucaaacgucuuccagacacgggcggggugccuuauuggagcagaacauguuaacaauuccuaugaaugcgauaucccgauuggagccgggaucugugcuagcuaucaaacccaaacaaauagccccagacgucgacguuccguggcuagucaaagcaucaucgccuacacuaugagucuuggggccgaaaauuccguugcuuacaguaacaacaguaucgcuauccccaccaauuuuacuauuaguguaacuacagagauucugccgguuuccaugacaaagacuuccguggauuguacgauguauauuugcggcgacagcacagagugcagcaaucugcugcugcaauacgguaguuucugcacccaauugaaccgugcucugacgggaauugcaguugagcaggacaagaauacucaagaaguauuugcacaagucaaacagauauacaagacgcccccgauuaaagauuucggcggguuuaacuuuagccaaauucuuccggaccccagcagacgccgccgaagcuuuauugaggaccugcuguuuaauaaaguuacccuugcugaugcugguuuuaucaagcaauacggagauugccugggagauaucgccgccagggauuugaucugugcgcaaaaguuuaacggccuuaccguucucccgccccuucugaccgaugaaaugauagcccaauacacuuccgcacuccuggcaggcacaauuacuuccggcuggacguuuggggccggggcagccuugcaaauuccguuugcuaugcaaauggcauaucguuucaaugguaucggcguaacacaaaauguccuuuaugagaaccagaaacucauugcuaaucaguuuaauuccgcuaucggcaagauucaagacagucucagcagcacggcgagcgcacuugguaaacuucaagacguugucaaccagaaugcucaagcccugaacacucugguaaaacaacuuagcucuaauuucggugcaauuagcuccgguccgaacgauauucugucacggcucccgaaagucgaagccgaaguccagaucgauaggcugaucacagggcgcuugcagagucuccaaaccuacgugacgcaacaacucauucgggcggcugaaauucgugcaagcgcuaaucuggccgcuaccaaaaugagugaguguguucucggucaaucaaagaggguugacuuuugcggcaaaggauaucauuugaugaguuuuccgcaaucugccccucaugggguaguauuucugcacguaacuuauguaccagcacaagaaaagaacuucaccacggccccagcaauaugccacgauggcaaagcucauuucccucgcgaaggggucuuuguaagcaauggaacccacugguuugucacacaacgcaacuuuuaugagccucaaaucauuacaaccgauaacacuuuugucuccgggaacugcgacguggugauuggaaucgucaacaacacugucuaugauccccugcaaccugaacuggauuccuuuaaagaagagcuugauaaguauuucaagaaccauaccagccccgacgucgauuugggagauauuagugggauuaaugcuagcguuguuaauauacaaaaggaaauagaucgauugaaugaaguggccaagaaucugaaugagucucugauugaccugcaggagcucggaaaguaugagcaauauauaaaauggcccuggucaggccgcaggcgucggcggcgcgguagcggcgguucaggaucuggguauauaccugaggccccacgagaugggcaggcuuauguacggaaagauggagaauggguguugcugaguacuuuccucggguaauaa (SEQ ID NO: 132)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGV

PT KLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDI

TEIYQAGSTPCNGVEGFNCYFPLQSYGFQP TNGVGYQPYRVVVLSFEL

HAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGIC ASYQTQTNSPRR

RSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPS

RSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTL VKQLSSNFGAISS

NDILSRL

KVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 133) WuS_3F_D2P_pVaxggatccgccaccatggactggacctggatactctttctcgtagcagcagccacacgagtgcattcaatgtttgtcttcttggtcctcttgccactggttagctcccagtgtgtgaatcttactacaaggacacaactgcccccagcttacacaaactcctttactaggggtgtatattacccagacaaagtgtttcgcagttctgtcctgcatagcacccaagaccttttccttccgttcttcagcaacgtcacctggttccatgctatccatgtcagtggaacgaatggcacaaagcggttcgataaccctgtcctgccctttaacgacggcgtctatttcgcttcaacagagaagagtaacattatcagaggatggatatttggtacaactcttgatagcaagacacaaagcctgctgattgtaaacaacgcgacaaatgtcgtcatcaaggtttgcgagtttcaattttgcaacgatcccttcttgggcgtgtactatcataagaacaataaaagctggatggagagcgaatttagggtgtatagctcagctaataactgtacatttgaatatgtctctcaacccttcctcatggaccttgagggaaagcaaggaaatttcaagaatctcagagaatttgtcttcaagaacatcgacgggtatttcaagatctactccaagcatacacccatcaacttggttagggaccttccgcaaggtttctcagcactggagcctctggtagatctccctattgggattaatattacaagatttcaaacactcctggccttgcatagatcctatcttacccctggggattccagctcaggttggaccgcgggtgccgcggcgtactatgtcggatatctccaacctcggacattcctgctgaaatacaatgaaaatgggaccatcactgatgccgttgattgtgctctcgatcctctgagtgagaccaaatgcactcttaagagttttacagtggagaaaggtatctatcaaactagtaatttcagagttcaaccaaccgagtcaatagtgcgttttccaaatatcactaatctgtgtccatttggggaagtcttcaatgctacccgattcgcaagtgtgtacgcctggaaccggaaacggatttctaactgcgttgccgattatagtgtcctctataattctgcttctttctctacttttaagtgctatggggtgtcccccaccaagctgaacgatctgtgtttcactaacgtctacgccgatagttttgtcattagaggggacgaggtacggcaaatcgcgcccggccaaacggggaaaattgccgattacaactacaagcttccagacgacttcacaggttgcgtgattgcatggaattctaataatctggacagtaaagtgggcggcaactataactatctttaccggctgtttcggaagagcaacttgaagcccttcgaacgcgacatatccaccgagatctatcaagccggaagtaccccgtgcaacggggtagaaggatttaattgttattttccattgcagtcttatggatttcagcccaccaatggtgtgggataccaaccttatagggttgttgttctctccttcgaactcctgcacgctccagctactgtatgtgggcctaagaaaagtactaatctcgttaagaataaatgcgtcaatttcaatttcaacggcttgaccgggactggagtgctcaccgaaagcaacaagaagtttctcccgtttcagcaattcggtagggatattgccgatacgacagatgcagtacgagatccccaaacactcgaaatcctggacattacgccatgtagctttggcggagtaagtgtcatcaccccagggactaacaccagtaaccaagttgcggtactctatcaggatgtgaactgcactgaggtacctgtagcaattcacgcagaccaattgacgccgacgtggcgcgtctatagtacaggaagtaacgtctttcagacaagagcgggttgtttgattggcgctgaacacgttaacaattcttacgagtgtgatatccccatcggtgcggggatctgcgccagctatcagacacaaaccaattccccacgaaggagacgttccgtggccagccagtcaataatcgcgtatactatgtctctgggtgcggagaattcagtggcctattccaataattctatagccattccaaccaattttactataagcgtcactacagagatcttgccagttagcatgacgaaaaccagcgtcgattgtaccatgtatatatgcggcgacagtaccgaatgctcaaatctgctgctccaatatggctcattttgcactcaacttaatagagctctgacagggatcgctgtcgaacaagataagaacactcaggaagttttcgcccaagttaagcagatatacaagaccccgcccatcaaggattttggcggatttaatttctctcagatcctgccggaccctagccgccgacgccggagctttatcgaagacttgctgtttaataaggttactctcgcagatgcaggcttcatcaagcaatacggtgactgccttggggatatcgctgctcgggacctgatctgtgctcagaaattcaacggtctcacggtgctgcccccactcctgaccgacgaaatgattgcccagtatacgtccgcattgctcgctggcaccatcactagcggctggacctttggggccggagccgcgctccaaataccttttgctatgcaaatggcttatcgcttcaatggtattggggttacgcaaaatgtcctctacgaaaatcaaaagctcatagctaaccaattcaatagcgctatagggaaaattcaagacagcctgagttccacagcaagcgccctcggcaaacttcaagatgtagtgaaccaaaatgctcaagcactcaatacactggtcaaacaactctcaagcaatttcggggcaatctcatctggtcctaatgacatattgagcaggctccccaaagtggaagcagaagtacaaatcgacaggctgattaccggacgactccaaagcttgcaaacttatgtaacccaacaacttatcagggctgcagaaatccgtgcaagcgctaacctcgccgctacgaagatgtcagaatgtgtacttgggcagtctaagagggttgatttctgtggaaaagggtaccatctgatgagttttccacagagcgctccacatggggtggtgtttctgcatgtaacctatgttcccgctcaagaaaagaattttactactgcccccgcaatttgccatgacgggaaagcccatttcccccgagagggagttttcgtgagtaacggaacgcactggtttgtcactcagagaaatttctacgagccccaaatcattacgaccgataatacattcgtaagcggtaactgcgatgtcgtcattggcatcgttaacaacactgtttatgatccccttcaacccgagcttgactcatttaaagaggaactggataagtactttaagaatcacacctctcccgatgtcgacctgggcgacatctctggaattaatgcctctgtcgtaaacatccaaaaggaaattgaccgactgaatgaggtggcaaagaatcttaatgaatccctgatcgatctgcaggagcttgggaagtatgagcaatacatcaaatggccatggtctggcagacggcgccggagaaggggctctggcggctctggaagcgggtatattccagaggcgcccagggatgggcaagcatatgttcggaaggatggggagtgggtgttgttgtccacgttccttggctagtga (SEQ ID NO: 134)ggauccgccaccauggacuggaccuggauacucuuucucguagcagcagccacacgagugcauucaauguuugucuucuugguccucuugccacugguuagcucccagugugugaaucuuacuacaaggacacaacugcccccagcuuacacaaacuccuuuacuagggguguauauuacccagacaaaguguuucgcaguucuguccugcauagcacccaagaccuuuuccuuccguucuucagcaacgucaccugguuccaugcuauccaugucaguggaacgaauggcacaaagcgguucgauaacccuguccugcccuuuaacgacggcgucuauuucgcuucaacagagaagaguaacauuaucagaggauggauauuugguacaacucuugauagcaagacacaaagccugcugauuguaaacaacgcgacaaaugucgucaucaagguuugcgaguuucaauuuugcaacgaucccuucuugggcguguacuaucauaagaacaauaaaagcuggauggagagcgaauuuaggguguauagcucagcuaauaacuguacauuugaauaugucucucaacccuuccucauggaccuugagggaaagcaaggaaauuucaagaaucucagagaauuugucuucaagaacaucgacggguauuucaagaucuacuccaagcauacacccaucaacuugguuagggaccuuccgcaagguuucucagcacuggagccucugguagaucucccuauugggauuaauauuacaagauuucaaacacuccuggccuugcauagauccuaucuuaccccuggggauuccagcucagguuggaccgcgggugccgcggcguacuaugucggauaucuccaaccucggacauuccugcugaaauacaaugaaaaugggaccaucacugaugccguugauugugcucucgauccucugagugagaccaaaugcacucuuaagaguuuuacaguggagaaagguaucuaucaaacuaguaauuucagaguucaaccaaccgagucaauagugcguuuuccaaauaucacuaaucuguguccauuuggggaagucuucaaugcuacccgauucgcaaguguguacgccuggaaccggaaacggauuucuaacugcguugccgauuauaguguccucuauaauucugcuucuuucucuacuuuuaagugcuauggggugucccccaccaagcugaacgaucuguguuucacuaacgucuacgccgauaguuuugucauuagaggggacgagguacggcaaaucgcgcccggccaaacggggaaaauugccgauuacaacuacaagcuuccagacgacuucacagguugcgugauugcauggaauucuaauaaucuggacaguaaagugggcggcaacuauaacuaucuuuaccggcuguuucggaagagcaacuugaagcccuucgaacgcgacauauccaccgagaucuaucaagccggaaguaccccgugcaacgggguagaaggauuuaauuguuauuuuccauugcagucuuauggauuucagcccaccaauggugugggauaccaaccuuauaggguuguuguucucuccuucgaacuccugcacgcuccagcuacuguaugugggccuaagaaaaguacuaaucucguuaagaauaaaugcgucaauuucaauuucaacggcuugaccgggacuggagugcucaccgaaagcaacaagaaguuucucccguuucagcaauucgguagggauauugccgauacgacagaugcaguacgagauccccaaacacucgaaauccuggacauuacgccauguagcuuuggcggaguaagugucaucaccccagggacuaacaccaguaaccaaguugcgguacucuaucaggaugugaacugcacugagguaccuguagcaauucacgcagaccaauugacgccgacguggcgcgucuauaguacaggaaguaacgucuuucagacaagagcggguuguuugauuggcgcugaacacguuaacaauucuuacgagugugauauccccaucggugcggggaucugcgccagcuaucagacacaaaccaauuccccacgaaggagacguuccguggccagccagucaauaaucgcguauacuaugucucugggugcggagaauucaguggccuauuccaauaauucuauagccauuccaaccaauuuuacuauaagcgucacuacagagaucuugccaguuagcaugacgaaaaccagcgucgauuguaccauguauauaugcggcgacaguaccgaaugcucaaaucugcugcuccaauauggcucauuuugcacucaacuuaauagagcucugacagggaucgcugucgaacaagauaagaacacucaggaaguuuucgcccaaguuaagcagauauacaagaccccgcccaucaaggauuuuggcggauuuaauuucucucagauccugccggacccuagccgccgacgccggagcuuuaucgaagacuugcuguuuaauaagguuacucucgcagaugcaggcuucaucaagcaauacggugacugccuuggggauaucgcugcucgggaccugaucugugcucagaaauucaacggucucacggugcugcccccacuccugaccgacgaaaugauugcccaguauacguccgcauugcucgcuggcaccaucacuagcggcuggaccuuuggggccggagccgcgcuccaaauaccuuuugcuaugcaaauggcuuaucgcuucaaugguauugggguuacgcaaaauguccucuacgaaaaucaaaagcucauagcuaaccaauucaauagcgcuauagggaaaauucaagacagccugaguuccacagcaagcgcccucggcaaacuucaagauguagugaaccaaaaugcucaagcacucaauacacuggucaaacaacucucaagcaauuucggggcaaucucaucugguccuaaugacauauugagcaggcuccccaaaguggaagcagaaguacaaaucgacaggcugauuaccggacgacuccaaagcuugcaaacuuauguaacccaacaacuuaucagggcugcagaaauccgugcaagcgcuaaccucgccgcuacgaagaugucagaauguguacuugggcagucuaagaggguugauuucuguggaaaaggguaccaucugaugaguuuuccacagagcgcuccacauggggugguguuucugcauguaaccuauguucccgcucaagaaaagaauuuuacuacugcccccgcaauuugccaugacgggaaagcccauuucccccgagagggaguuuucgugaguaacggaacgcacugguuugucacucagagaaauuucuacgagccccaaaucauuacgaccgauaauacauucguaagcgguaacugcgaugucgucauuggcaucguuaacaacacuguuuaugauccccuucaacccgagcuugacucauuuaaagaggaacuggauaaguacuuuaagaaucacaccucucccgaugucgaccugggcgacaucucuggaauuaaugccucugucguaaacauccaaaaggaaauugaccgacugaaugagguggcaaagaaucuuaaugaaucccugaucgaucugcaggagcuugggaaguaugagcaauacaucaaauggccauggucuggcagacggcgccggagaaggggcucuggcggcucuggaagcggguauauuccagaggcgcccagggaugggcaagcauauguucggaaggauggggaguggguguuguuguccacguuccuuggcuaguga (SEQ ID NO: 135)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSGPNDILSRLPKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 136) WuS_3F_2P_GlyDSol_pVaxggatccgccaccatggactggacatggattttgtttcttgtggcggctgcaacgcgagttcattccatgtttgttttcctggttctcttgccgctcgtctctagtcaatgcgtcaatctgacgacaagaacgcaacttccccctgcctacaccaatagttttacccgtggcgtctattatccagataaagtgtttcgaagttcctgccttcattccacgcaagatctcttccttccattcttctctaatgtcacctggtttcatgcgatccacgtgtctggaaccaacgggactaaacgattcgacaatcccgtcctgccatttaacgatggagtatacttcgcatccaccgagaaatctggcattataagagggtggatattcgggactacactcgacagcaagacacaaagtctcctgattgttaacaacgcgacaaacgtcgtaattaaagtttgcgaatttcaattttgtaatgacccgttcttgggcgtgtattatcacaagaataacaaaagttggatggaatccgaattccgggtttattccagcgcgaataattgcacatttgaatacgttagccaacctttcctgatggatctcgaaggcaaacaaggaaactttaagaatcttcgggaattcgttttcaagaacatcgacgggtactttaagatatactcaaaacacaccccaattaatttggtccgagatctgccgcaaggctttagtgcgcttgagcccctcgtagatctcccaattggtattaatatcacacgctttcaaaccctgctggcattgcatcggagctatctgactcccggcgatagttcttcagggtggacggcgggtgccgctgcttactatgtaggctatctgcagcctcgtacatttctcctcaaatacaatgaaaatggcactattaccgacgccgttgattgcgctctggacccactgagcgagactaaatgcaccctcaaatcattcactgttgagaagggaatttaccaaacatcaaacttcagggtccaacctacggaaagcatcgtgcggttccccaacatcactaacctctgcccctttggagaagtatttaacgctacaaggttcgcttccgtctacgcctggaacaggaaaagaatcagtaattgcgtggctgattactccgtgctgtacaattccgcctcattttctacatttaagtgttatggcgttaacgggactaagcttaacgacctctgcttcacaaacgtctatgccgacagctttgtcattcgcggggatgaagtaagacagatagcacccggtcaaactggcaaaattgctgattacaattacaagttgccagatgatttcactggatgcgttatagcatggaactctaacaaccttgactcaaaggttggtggcaactataattatttgtatcgcctgtttcgcaaatctaatctcaagcctttcgagcgcgacataaatacgaccatataccaagcggggtccaccccttgtaatggagtcgaggggtttaattgctattttccgttgcaatcctacgggttccaaccaacaaacggcgtcggctatcaaccctatcgggttgtcgtactctcattcgagctcaaccatgcaccagcaacagtttgtggccccaagaagagcacaaatttggtcaagaataaatgcgttaattttaatttcaatggtctgactggcacaggggttcttaccgaatcaaataagaagtttctgccatttcagcagttcggaagggactgtgcagggaccacagatgccgttagagacccccaaacactcgaaattctggacatcacgccatgcagtttcggtggtgttagcgtgattactccgggtactaatacgtccaaccaagtggctgtgttgtatcaagacgttaactgtaccgaagttcctgtagcaatccatgccggtcaactgacccccacgtggcgagtttatagcaccggttccaacgtctttcaaacaagagccggatgtctcataggcgctgaacatgtgaataattcatacgaatgtgacattccaatcggcgcagggatttgcgcctcatatcagacacaaactaactccccgagaagacgtcgctcagtggcgtcacaaagcatcatcgcttatacgatgagcctctgcgccgagaactctgtcgcatattctaacaactctattgcaattcctacaaattttacaatttcttgcactactgagatcctgcccgtaagcatgacgaaaacatcctgcgactgcacaatgtatatctgtggcgactcaactgagtgctccaatctcctcttgcaatacggatctttctgtactcaactcaacagagcacttacaggaatagccgtcgaacaagacaagaacacacaagaggtcttcgcccaagtaaagcaatgttacaaaaccccacctattaaagactttggtgggtttaatttctcacagattcttccagatccttcccgtagaaggagaagctttattgaagacctcttgtttaataaagtcactcttgcagacgctgggtttattaaacaatatggagactgcttgggagacatagcggcaagagacctgatctgcgctcaaaagtttaatgggtgcactgtgttgccaccccttctgaccgacgagatgatcgctcagtataccagtgccttgctggcagggaccataactagcggatggactttcggtgcaggagctgctctgcaaatcccttttgcgatgcaaatggcctacaggtttaatggtataggagttactcagaatgtcctgtacgaaaatcaaaagctgatcgccaatcaattcaacagtgctattgggaaaatacaggacagtttgagttcaacagcgagcgctctcggcaaactgcaggatgttgtgaatcaaaacgcgcaagctttgaacactcttgtgaagcagctttcatccaacttcggagcgatctcatccgtcctgaacgacatattgtcaagacttgacccacctgaagcggaagttcagatagaccgactcataacgggccgacttcagtccttgcagacatacgtgacccaacaacttatccgcgcagccgaaataagggcttcagctaaccttgcagcaaccaaaatgtcagagtgcgtgctcggtcaaagcaagcgggtagacttttgtggcaaggggtatcatcttatgtcctttcctcaatccgcccctcacggggtggtcttcttgcactgcacttatgtacctgctcaagagaagaattttacgaccgcccctgcgatctgtcacgacgggaaagcacatttcccccgcgagggagtctttgtgtctaatggtactcattggtttgttacgcagcggaacttttacgaacctcaaataattacaacggataatacagatgttagtgggaattgcgacgtggtgatcggtatagtcaacaatacggtgtatgatccacttcaaccagaacttgattcctttaaggaagagctggacaaatatttcaagaaccatacatcccctgacgtggaccttggcgatataagcggcattaatgcttcagtggtcaatatacaaaaggaaatcgatcgcctgaatgaggtcgcaaagaatttgaatgagtccctgatcgacctgcaagagctcgggaaatatgagcagtacatcaagtggccctggtcaggtagacgtaggcggcgccggggcagtggcggctcagggagcggttatatacccgaagcccctagagatgggcaagcttatgtccgaaaggacggcgaatgggtgctcctttccactttcttgggataatag (SEQ ID NO: 137)ggauccgccaccauggacuggacauggauuuuguuucuuguggcggcugcaacgcgaguucauuccauguuuguuuuccugguucucuugccgcucgucucuagucaaugcgucaaucugacgacaagaacgcaacuucccccugccuacaccaauaguuuuacccguggcgucuauuauccagauaaaguguuucgaaguuccugccuucauuccacgcaagaucucuuccuuccauucuucucuaaugucaccugguuucaugcgauccacgugucuggaaccaacgggacuaaacgauucgacaaucccguccugccauuuaacgauggaguauacuucgcauccaccgagaaaucuggcauuauaagaggguggauauucgggacuacacucgacagcaagacacaaagucuccugauuguuaacaacgcgacaaacgucguaauuaaaguuugcgaauuucaauuuuguaaugacccguucuugggcguguauuaucacaagaauaacaaaaguuggauggaauccgaauuccggguuuauuccagcgcgaauaauugcacauuugaauacguuagccaaccuuuccugauggaucucgaaggcaaacaaggaaacuuuaagaaucuucgggaauucguuuucaagaacaucgacggguacuuuaagauauacucaaaacacaccccaauuaauuugguccgagaucugccgcaaggcuuuagugcgcuugagccccucguagaucucccaauugguauuaauaucacacgcuuucaaacccugcuggcauugcaucggagcuaucugacucccggcgauaguucuucaggguggacggcgggugccgcugcuuacuauguaggcuaucugcagccucguacauuucuccucaaauacaaugaaaauggcacuauuaccgacgccguugauugcgcucuggacccacugagcgagacuaaaugcacccucaaaucauucacuguugagaagggaauuuaccaaacaucaaacuucaggguccaaccuacggaaagcaucgugcgguuccccaacaucacuaaccucugccccuuuggagaaguauuuaacgcuacaagguucgcuuccgucuacgccuggaacaggaaaagaaucaguaauugcguggcugauuacuccgugcuguacaauuccgccucauuuucuacauuuaaguguuauggcguuaacgggacuaagcuuaacgaccucugcuucacaaacgucuaugccgacagcuuugucauucgcggggaugaaguaagacagauagcacccggucaaacuggcaaaauugcugauuacaauuacaaguugccagaugauuucacuggaugcguuauagcauggaacucuaacaaccuugacucaaagguugguggcaacuauaauuauuuguaucgccuguuucgcaaaucuaaucucaagccuuucgagcgcgacauaaauacgaccauauaccaagcgggguccaccccuuguaauggagucgagggguuuaauugcuauuuuccguugcaauccuacggguuccaaccaacaaacggcgucggcuaucaacccuaucggguugucguacucucauucgagcucaaccaugcaccagcaacaguuuguggccccaagaagagcacaaauuuggucaagaauaaaugcguuaauuuuaauuucaauggucugacuggcacagggguucuuaccgaaucaaauaagaaguuucugccauuucagcaguucggaagggacugugcagggaccacagaugccguuagagacccccaaacacucgaaauucuggacaucacgccaugcaguuucggugguguuagcgugauuacuccggguacuaauacguccaaccaaguggcuguguuguaucaagacguuaacuguaccgaaguuccuguagcaauccaugccggucaacugacccccacguggcgaguuuauagcaccgguuccaacgucuuucaaacaagagccggaugucucauaggcgcugaacaugugaauaauucauacgaaugugacauuccaaucggcgcagggauuugcgccucauaucagacacaaacuaacuccccgagaagacgucgcucaguggcgucacaaagcaucaucgcuuauacgaugagccucugcgccgagaacucugucgcauauucuaacaacucuauugcaauuccuacaaauuuuacaauuucuugcacuacugagauccugcccguaagcaugacgaaaacauccugcgacugcacaauguauaucuguggcgacucaacugagugcuccaaucuccucuugcaauacggaucuuucuguacucaacucaacagagcacuuacaggaauagccgucgaacaagacaagaacacacaagaggucuucgcccaaguaaagcaauguuacaaaaccccaccuauuaaagacuuugguggguuuaauuucucacagauucuuccagauccuucccguagaaggagaagcuuuauugaagaccucuuguuuaauaaagucacucuugcagacgcuggguuuauuaaacaauauggagacugcuugggagacauagcggcaagagaccugaucugcgcucaaaaguuuaaugggugcacuguguugccaccccuucugaccgacgagaugaucgcucaguauaccagugccuugcuggcagggaccauaacuagcggauggacuuucggugcaggagcugcucugcaaaucccuuuugcgaugcaaauggccuacagguuuaaugguauaggaguuacucagaauguccuguacgaaaaucaaaagcugaucgccaaucaauucaacagugcuauugggaaaauacaggacaguuugaguucaacagcgagcgcucucggcaaacugcaggauguugugaaucaaaacgcgcaagcuuugaacacucuugugaagcagcuuucauccaacuucggagcgaucucauccguccugaacgacauauugucaagacuugacccaccugaagcggaaguucagauagaccgacucauaacgggccgacuucaguccuugcagacauacgugacccaacaacuuauccgcgcagccgaaauaagggcuucagcuaaccuugcagcaaccaaaaugucagagugcgugcucggucaaagcaagcggguagacuuuuguggcaagggguaucaucuuauguccuuuccucaauccgccccucacgggguggucuucuugcacugcacuuauguaccugcucaagagaagaauuuuacgaccgccccugcgaucugucacgacgggaaagcacauuucccccgcgagggagucuuugugucuaaugguacucauugguuuguuacgcagcggaacuuuuacgaaccucaaauaauuacaacggauaauacagauguuagugggaauugcgacguggugaucgguauagucaacaauacgguguaugauccacuucaaccagaacuugauuccuuuaaggaagagcuggacaaauauuucaagaaccauacauccccugacguggaccuuggcgauauaagcggcauuaaugcuucaguggucaauauacaaaaggaaaucgaucgccugaaugaggucgcaaagaauuugaaugagucccugaucgaccugcaagagcucgggaaauaugagcaguacaucaaguggcccuggucagguagacguaggcggcgccggggcaguggcggcucagggagcgguuauauacccgaagccccuagagaugggcaagcuuauguccgaaaggacggcgaaugggugcuccuuuccacuuucuugggauaauag (SEQ ID NO: 138)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSCLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSGIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVNGTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDINTTIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELNHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDCAGTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHAGQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLCAENSVAYSNNSIAIPTNFTISCTTEILPVSMTKTSCDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQCYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGCTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHCTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTDVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 139) WuS_3F_2P_Gly_pVaxggatccgccaccatggattggacctggatacttttcctcgtggccgcagcaacaagagtccactccatgttcgtcttcttggtacttttgccactcgtcagttctcagtgcgttaacctgactactagaacccaattgcccccggcatacacaaactctttcacccggggtgtctactatcccgacaaagtgtttagaagtagcgtgctgcacagcacccaagatctctttctgccattcttctcaaacgtcacctggtttcacgccatccatgtaagcgggaccaacggcacaaagcgttttgataaccctgttttgccattcaatgatggcgtgtattttgcttccactgagaaaagcaacatcattagagggtggatatttggcacaacgcttgactccaagacgcagagtcttttgatagtaaacaacgcaactaatgtggtcattaaagtctgtgaatttcaattttgcaatgaccccttccttggagtctattaccacaagaacaacaaaagctggatggaaagcgaatttagggtctacagctctgccaataactgcacattcgaatacgtcagccaaccattcttgatggacctggaaggcaagcaaggaaactttaagaatctgagggaatttgtgtttaagaatatcgacggatattttaagatctattccaagcatactcccattaatctcgttcgtgaccttcctcagggtttctctgcattggaacccctcgtagatttgcccattgggattaatatcactagattccagacgctgcttgcactccatcgatcttatctgacccctggtgactcctcttccgggtggacggcgggtgctgcagcctactacgttggctatttgcaacctaggacctttctgttgaagtataatgagaatgggactattactgatgccgttgattgcgccctcgatccgctgtcagaaacaaagtgcaccctgaagagcttcacagtagaaaagggaatctatcaaacctcaaatttccgcgttcaaccaactgaatcaatcgtgcgttttcctaacatcacaaatctgtgtccgtttggagaagtatttaatgcgacgcgtttcgcaagcgtctacgcgtggaatcgcaaacgtatctctaattgcgtagcagattattctgtgctgtacaatagcgcatctttctcaacgtttaagtgctacggcgttaatgggaccaagctgaatgatctctgtttcactaatgtgtacgcagacagttttgtaattagaggagacgaggttaggcaaatagcaccgggtcaaactggcaaaatcgccgactataactacaagctccctgatgacttcacgggctgcgtaattgcttggaactctaataacctggactctaaagtcggcgggaattataattatctctatcggttgtttcgaaaatccaatctcaaaccctttgagcgggacatcaatactacaatttatcaagctggtagtactccttgcaatggggtagaaggcttcaattgttatttcccccttcaatcttacggatttcaacccacgaacggcgtagggtaccagccctatcgagtggtggtactgtcattcgaacttaatcacgccccagcaacagtctgcgggcctaagaaaagcacgaatcttgtcaagaataagtgtgtaaatttcaacttcaatggtcttacaggcacgggagtgctcactgagtctaataagaaatttcttcctttccaacaattcggtcgtgatattgccgatactactgatgcagtccgagatccacaaactctcgaaatcctcgatattactccttgtagttttggcggcgtctccgtgatcaccccagggaccaacactagtaaccaagtggcggtgctctaccaagatgttaactgcacagaagtcccggtagcgatccatgccgaccagctcactcccacatggcgtgtttacagcacagggtcaaacgttttccagacccgtgccggatgtcttataggagccgaacacgtaaataacagttatgaatgcgatatcccaattggtgcaggtatctgtgcgtcatatcaaacccaaactaattctccgagacgacgacggagcgttgcctcacaatcaataatcgcctacacaatgtccctcggtgccgaaaattcagtcgcttactctaacaatagcattgctatccctaccaacttcactatttctgttaccacggaaattttgcctgtatccatgaccaaaacatctgttgattgcacgatgtacatctgcggggattctaccgaatgttctaacctgcttctgcaatacggctccttctgcacccaattgaaccgcgcactgactgggattgctgtggaacaagacaagaatactcaagaagtatttgcccaggtcaaacagatttacaaaactcccccaattaaagatttcggcggtttcaattttagtcaaattctgccagatccaagtcgacgccgcaggagctttattgaggacctgctctttaataaagtcacgctggccgacgccggcttcataaaacagtatggcgattgtcttggagacatcgccgcccgcgacctcatttgcgcacaaaagttcaatgggctcaccgtgttgccaccactgctcacagatgagatgatcgcacagtacacgagcgcccttcttgccggcactatcacgtctggttggacgttcggtgccggagccgctctgcaaattccctttgcaatgcaaatggcctatagatttaatggaattggcgtaacacagaacgtgttgtacgagaaccagaagctcattgccaaccagttcaattccgctattggcaaaatacaagactctctcagctcaactgctagcgcactgggaaaattgcaagacgtagtcaatcaaaatgcccaagccctcaatactctcgtcaaacagttgtcttccaactttggggctatcagtagtgtactcaatgacattctttcaagactggacccgcccgaggcggaagtccaaattgatcgtctgataactggaaggttgcaaagccttcagacctacgttacgcaacaacttattagggctgccgaaataagggcatccgctaatctggcagctacaaagatgtctgaatgtgttttgggacagagcaaacgggttgacttctgcggtaaaggttaccatctcatgtcttttccacaaagcgcaccgcacggagtcgtcttcctgcatgtaacatacgtcccagcccaagaaaagaattttaccacagccccagccatctgccacgacggcaaggcgcatttcccaagggaaggcgtgtttgtatccaacgggacgcattggtttgtcactcaaaggaacttttacgaaccccaaattattaccactgataacaccttcgtttctgggaactgtgatgtcgtgattgggatagtaaacaacacggtatatgatccactgcaaccagaactggattccttcaaagaagagctggacaaatacttcaagaatcatactagtcctgacgtcgacctgggcgatatcagtggaatcaacgctagcgtcgtaaacattcaaaaggagatcgatagacttaacgaggtcgccaagaatctcaatgaaagcctcatcgatttgcaagaactcggaaaatatgagcaatacataaaatggccatggtctggcaggagaagacgcaggagaggtagcggcggcagcggatcagggtacattccggaagcccccagggacggacaggcatatgtccgcaaggacggagaatgggttcttcttagcacttttctggggtaataa (SEQ ID NO: 140)ggauccgccaccauggauuggaccuggauacuuuuccucguggccgcagcaacaagaguccacuccauguucgucuucuugguacuuuugccacucgucaguucucagugcguuaaccugacuacuagaacccaauugcccccggcauacacaaacucuuucacccggggugucuacuaucccgacaaaguguuuagaaguagcgugcugcacagcacccaagaucucuuucugccauucuucucaaacgucaccugguuucacgccauccauguaagcgggaccaacggcacaaagcguuuugauaacccuguuuugccauucaaugauggcguguauuuugcuuccacugagaaaagcaacaucauuagaggguggauauuuggcacaacgcuugacuccaagacgcagagucuuuugauaguaaacaacgcaacuaauguggucauuaaagucugugaauuucaauuuugcaaugaccccuuccuuggagucuauuaccacaagaacaacaaaagcuggauggaaagcgaauuuagggucuacagcucugccaauaacugcacauucgaauacgucagccaaccauucuugauggaccuggaaggcaagcaaggaaacuuuaagaaucugagggaauuuguguuuaagaauaucgacggauauuuuaagaucuauuccaagcauacucccauuaaucucguucgugaccuuccucaggguuucucugcauuggaaccccucguagauuugcccauugggauuaauaucacuagauuccagacgcugcuugcacuccaucgaucuuaucugaccccuggugacuccucuuccggguggacggcgggugcugcagccuacuacguuggcuauuugcaaccuaggaccuuucuguugaaguauaaugagaaugggacuauuacugaugccguugauugcgcccucgauccgcugucagaaacaaagugcacccugaagagcuucacaguagaaaagggaaucuaucaaaccucaaauuuccgcguucaaccaacugaaucaaucgugcguuuuccuaacaucacaaaucuguguccguuuggagaaguauuuaaugcgacgcguuucgcaagcgucuacgcguggaaucgcaaacguaucucuaauugcguagcagauuauucugugcuguacaauagcgcaucuuucucaacguuuaagugcuacggcguuaaugggaccaagcugaaugaucucuguuucacuaauguguacgcagacaguuuuguaauuagaggagacgagguuaggcaaauagcaccgggucaaacuggcaaaaucgccgacuauaacuacaagcucccugaugacuucacgggcugcguaauugcuuggaacucuaauaaccuggacucuaaagucggcgggaauuauaauuaucucuaucgguuguuucgaaaauccaaucucaaacccuuugagcgggacaucaauacuacaauuuaucaagcugguaguacuccuugcaaugggguagaaggcuucaauuguuauuucccccuucaaucuuacggauuucaacccacgaacggcguaggguaccagcccuaucgaguggugguacugucauucgaacuuaaucacgccccagcaacagucugcgggccuaagaaaagcacgaaucuugucaagaauaaguguguaaauuucaacuucaauggucuuacaggcacgggagugcucacugagucuaauaagaaauuucuuccuuuccaacaauucggucgugauauugccgauacuacugaugcaguccgagauccacaaacucucgaaauccucgauauuacuccuuguaguuuuggcggcgucuccgugaucaccccagggaccaacacuaguaaccaaguggcggugcucuaccaagauguuaacugcacagaagucccgguagcgauccaugccgaccagcucacucccacauggcguguuuacagcacagggucaaacguuuuccagacccgugccggaugucuuauaggagccgaacacguaaauaacaguuaugaaugcgauaucccaauuggugcagguaucugugcgucauaucaaacccaaacuaauucuccgagacgacgacggagcguugccucacaaucaauaaucgccuacacaaugucccucggugccgaaaauucagucgcuuacucuaacaauagcauugcuaucccuaccaacuucacuauuucuguuaccacggaaauuuugccuguauccaugaccaaaacaucuguugauugcacgauguacaucugcggggauucuaccgaauguucuaaccugcuucugcaauacggcuccuucugcacccaauugaaccgcgcacugacugggauugcuguggaacaagacaagaauacucaagaaguauuugcccaggucaaacagauuuacaaaacucccccaauuaaagauuucggcgguuucaauuuuagucaaauucugccagauccaagucgacgccgcaggagcuuuauugaggaccugcucuuuaauaaagucacgcuggccgacgccggcuucauaaaacaguauggcgauugucuuggagacaucgccgcccgcgaccucauuugcgcacaaaaguucaaugggcucaccguguugccaccacugcucacagaugagaugaucgcacaguacacgagcgcccuucuugccggcacuaucacgucugguuggacguucggugccggagccgcucugcaaauucccuuugcaaugcaaauggccuauagauuuaauggaauuggcguaacacagaacguguuguacgagaaccagaagcucauugccaaccaguucaauuccgcuauuggcaaaauacaagacucucucagcucaacugcuagcgcacugggaaaauugcaagacguagucaaucaaaaugcccaagcccucaauacucucgucaaacaguugucuuccaacuuuggggcuaucaguaguguacucaaugacauucuuucaagacuggacccgcccgaggcggaaguccaaauugaucgucugauaacuggaagguugcaaagccuucagaccuacguuacgcaacaacuuauuagggcugccgaaauaagggcauccgcuaaucuggcagcuacaaagaugucugaauguguuuugggacagagcaaacggguugacuucugcgguaaagguuaccaucucaugucuuuuccacaaagcgcaccgcacggagucgucuuccugcauguaacauacgucccagcccaagaaaagaauuuuaccacagccccagccaucugccacgacggcaaggcgcauuucccaagggaaggcguguuuguauccaacgggacgcauugguuugucacucaaaggaacuuuuacgaaccccaaauuauuaccacugauaacaccuucguuucugggaacugugaugucgugauugggauaguaaacaacacgguauaugauccacugcaaccagaacuggauuccuucaaagaagagcuggacaaauacuucaagaaucauacuaguccugacgucgaccugggcgauaucaguggaaucaacgcuagcgucguaaacauucaaaaggagaucgauagacuuaacgaggucgccaagaaucucaaugaaagccucaucgauuugcaagaacucggaaaauaugagcaauacauaaaauggccauggucuggcaggagaagacgcaggagagguagcggcggcagcggaucaggguacauuccggaagcccccagggacggacaggcauauguccgcaaggacggagaauggguucuucuuagcacuuuucugggguaauaa (SEQ ID NO: 141)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVNGTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDINTTIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELNHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 142) WuS_3F_2P_NoClev_pVaxggatccgccaccatggattggacgtggattctgtttctggtggccgcagcgacaagggtgcattcaatgtttgtgttcctggtcctgctgccactggtctcatcacagtgtgtaaacctgactacaagaacgcagcttccgcctgcctacacgaacagcttcaccaggggagtgtattatcctgacaaagtctttaggagctctgttctccactccactcaagacctgtttctgcccttcttcagtaacgtgacttggtttcacgcaatacatgtctccggcacaaatggaaccaaaagattcgataaccctgttctcccattcaatgatggagtatattttgctagcactgaaaagtctaacattattagaggctggatatttggcacgacattggactccaagacgcaaagtctcttgattgtgaacaacgcaacaaacgtggtgataaaagtttgcgaattccaattttgcaatgacccattcctgggagtttactaccacaagaataacaaaagctggatggaatccgagttccgggtttactcctctgctaacaactgtacctttgagtatgtgagtcaaccattccttatggatctcgaaggaaaacaaggtaacttcaagaacctgagggagtttgtgtttaagaatatcgatggctattttaagatttatagcaaacacactccgattaatctggtgagagatctcccgcaaggattttctgctttggagccattggttgacttgcctattggaatcaacatcacccgttttcaaactctgcttgcgctgcatagatcctaccttacgcctggcgattcaagcagtggctggaccgcgggagcggccgcctattatgtaggctacttgcagcctcgcacctttctcctcaagtacaatgaaaacggcacaattacagacgcagtggattgtgctctggaccccctcagtgaaactaaatgtaccctgaaaagcttcactgttgagaaaggcatatatcaaacctcaaactttagagtgcaacctactgaaagcattgtaagattccctaacattacaaacctgtgcccctttggcgaagtctttaatgcaacccggtttgctagcgtgtatgcttggaaccgcaagaggatatccaattgcgtcgcagattattccgtcctgtataactctgccagctttagtaccttcaaatgttatggggtatctcccacaaaactcaatgatctttgtttcacaaatgtctatgctgactcctttgttatcagaggggacgaagttcgccaaattgctccaggtcaaacaggaaagattgcagattataactacaagcttcccgacgattttacaggttgtgtgatagcttggaactccaataatctggattccaaggtaggcgggaactacaattatctctacaggctcttccggaaatccaatctcaagccgttcgaaagggatataagcactgagatctatcaagcaggcagtacaccctgtaacggagtagagggcttcaactgctactttccactgcaatcctatgggtttcaaccgactaacggtgtcgggtaccaaccctatcgtgtcgtggtcctgtcctttgagcttctgcacgctcctgctaccgtttgcggccccaagaaaagcacgaatttggtcaagaacaaatgtgtcaactttaacttcaacggattgacagggaccggagtattgaccgaatctaataagaaatttctgcccttccaacaattcggacgggacatagcagacacaaccgatgctgtcagggacccacagacacttgaaatactcgatatcaccccatgcagctttggcggagtctcagtcattacgcctggcaccaatacttccaatcaagttgcagtgctctatcaggatgtcaattgtactgaggtccccgtcgccatccacgcggatcaacttacccccacatggcgagtatatagtaccgggagcaacgtctttcaaacccgagcaggatgtctgataggtgccgaacacgtaaacaacagctacgaatgtgatatcccgatcggcgcagggatttgcgctagctaccaaacccaaactaattctccgcgccgccgcaggtccgtagcaagtcaatcaataatagcatacaccatgtcattgggagctgaaaacagcgtggcatatagcaacaattccatagctatccctacaaatttcacgatttctgttaccaccgaaattctgccagtgagcatgaccaaaacctcagtggattgtacgatgtacatatgcggcgattccacggaatgttccaatctccttttgcaatacggcagcttttgtacccaactgaatagagctctgacgggtatagcagtagagcaggataagaacactcaagaggtgtttgcccaggtcaaacaaatttacaagactcccccaataaaagactttggcggcttcaatttcagccaaatcttgccagacccttccaggcggcggcgctcatttatcgaagatttgcttttcaataaagtcaccctggccgacgccggatttattaaacaatacggcgattgtctgggcgacatcgccgcaagggacctcatctgtgcgcaaaagttcaatggcctgacggtgcttccaccactcctgactgatgagatgattgcccaatacacatctgccctgctggctggtacaataacgagtgggtggacctttggggctggagcagcattgcaaattccattcgccatgcaaatggcatatcgttttaacggcattggagtgactcaaaatgtgctgtatgaaaaccaaaagcttattgcaaatcagtttaattccgccattggcaaaatccaggatagcctcagtagtacagcaagcgccttggggaaactgcaagatgtggttaatcaaaatgcacaagctctcaataccctggtcaagcaacttagtagtaactttggtgccatcagcagcgttctcaacgacatcctgagtcgtcttgatcccccagaggcagaggttcaaattgaccggcttatcactggaaggcttcaatccctgcaaacttacgtgactcagcaactgatacgcgctgcagaaattcgggcctcagcaaaccttgccgcgacaaagatgagcgaatgcgtgctgggacaatccaagcgggtcgacttttgtggtaaaggctatcatctgatgagcttcccacagtccgctccacacggcgtcgttttcctgcacgtgacctatgtgccagcacaggagaagaactttacaacagccccggctatctgccacgatggcaaagctcactttcctagagagggagtgtttgtaagcaatggaacccattggttcgttacacaaagaaacttttatgagccgcaaattatcacaacagataatacattcgtctccgggaactgtgacgttgtgatagggattgtcaacaacacagtgtacgaccccctgcaacccgagctggattcatttaaagaagaactcgacaagtacttcaagaatcatactagtccagatgtggatctgggcgatatatcaggaatcaatgccagcgtggtcaatattcaaaaggagattgatagactgaacgaggttgccaagaatctgaatgaaagcctgatcgatctgcaagaattgggcaagtatgagcagtacattaaatggccctggtctggcgggagcggcggatctgggtctggatatattcccgaagctcctagagatggacaagcttacgtccgtaaagacggcgagtgggttcttctctccacattcctcggctgatga (SEQ ID NO: 143)ggauccgccaccauggauuggacguggauucuguuucugguggccgcagcgacaagggugcauucaauguuuguguuccugguccugcugccacuggucucaucacaguguguaaaccugacuacaagaacgcagcuuccgccugccuacacgaacagcuucaccaggggaguguauuauccugacaaagucuuuaggagcucuguucuccacuccacucaagaccuguuucugcccuucuucaguaacgugacuugguuucacgcaauacaugucuccggcacaaauggaaccaaaagauucgauaacccuguucucccauucaaugauggaguauauuuugcuagcacugaaaagucuaacauuauuagaggcuggauauuuggcacgacauuggacuccaagacgcaaagucucuugauugugaacaacgcaacaaacguggugauaaaaguuugcgaauuccaauuuugcaaugacccauuccugggaguuuacuaccacaagaauaacaaaagcuggauggaauccgaguuccggguuuacuccucugcuaacaacuguaccuuugaguaugugagucaaccauuccuuauggaucucgaaggaaaacaagguaacuucaagaaccugagggaguuuguguuuaagaauaucgauggcuauuuuaagauuuauagcaaacacacuccgauuaaucuggugagagaucucccgcaaggauuuucugcuuuggagccauugguugacuugccuauuggaaucaacaucacccguuuucaaacucugcuugcgcugcauagauccuaccuuacgccuggcgauucaagcaguggcuggaccgcgggagcggccgccuauuauguaggcuacuugcagccucgcaccuuucuccucaaguacaaugaaaacggcacaauuacagacgcaguggauugugcucuggacccccucagugaaacuaaauguacccugaaaagcuucacuguugagaaaggcauauaucaaaccucaaacuuuagagugcaaccuacugaaagcauuguaagauucccuaacauuacaaaccugugccccuuuggcgaagucuuuaaugcaacccgguuugcuagcguguaugcuuggaaccgcaagaggauauccaauugcgucgcagauuauuccguccuguauaacucugccagcuuuaguaccuucaaauguuaugggguaucucccacaaaacucaaugaucuuuguuucacaaaugucuaugcugacuccuuuguuaucagaggggacgaaguucgccaaauugcuccaggucaaacaggaaagauugcagauuauaacuacaagcuucccgacgauuuuacagguugugugauagcuuggaacuccaauaaucuggauuccaagguaggcgggaacuacaauuaucucuacaggcucuuccggaaauccaaucucaagccguucgaaagggauauaagcacugagaucuaucaagcaggcaguacacccuguaacggaguagagggcuucaacugcuacuuuccacugcaauccuauggguuucaaccgacuaacggugucggguaccaacccuaucgugucgugguccuguccuuugagcuucugcacgcuccugcuaccguuugcggccccaagaaaagcacgaauuuggucaagaacaaaugugucaacuuuaacuucaacggauugacagggaccggaguauugaccgaaucuaauaagaaauuucugcccuuccaacaauucggacgggacauagcagacacaaccgaugcugucagggacccacagacacuugaaauacucgauaucaccccaugcagcuuuggcggagucucagucauuacgccuggcaccaauacuuccaaucaaguugcagugcucuaucaggaugucaauuguacugagguccccgucgccauccacgcggaucaacuuacccccacauggcgaguauauaguaccgggagcaacgucuuucaaacccgagcaggaugucugauaggugccgaacacguaaacaacagcuacgaaugugauaucccgaucggcgcagggauuugcgcuagcuaccaaacccaaacuaauucuccgcgccgccgcagguccguagcaagucaaucaauaauagcauacaccaugucauugggagcugaaaacagcguggcauauagcaacaauuccauagcuaucccuacaaauuucacgauuucuguuaccaccgaaauucugccagugagcaugaccaaaaccucaguggauuguacgauguacauaugcggcgauuccacggaauguuccaaucuccuuuugcaauacggcagcuuuuguacccaacugaauagagcucugacggguauagcaguagagcaggauaagaacacucaagagguguuugcccaggucaaacaaauuuacaagacucccccaauaaaagacuuuggcggcuucaauuucagccaaaucuugccagacccuuccaggcggcggcgcucauuuaucgaagauuugcuuuucaauaaagucacccuggccgacgccggauuuauuaaacaauacggcgauugucugggcgacaucgccgcaagggaccucaucugugcgcaaaaguucaauggccugacggugcuuccaccacuccugacugaugagaugauugcccaauacacaucugcccugcuggcugguacaauaacgaguggguggaccuuuggggcuggagcagcauugcaaauuccauucgccaugcaaauggcauaucguuuuaacggcauuggagugacucaaaaugugcuguaugaaaaccaaaagcuuauugcaaaucaguuuaauuccgccauuggcaaaauccaggauagccucaguaguacagcaagcgccuuggggaaacugcaagaugugguuaaucaaaaugcacaagcucucaauacccuggucaagcaacuuaguaguaacuuuggugccaucagcagcguucucaacgacauccugagucgucuugaucccccagaggcagagguucaaauugaccggcuuaucacuggaaggcuucaaucccugcaaacuuacgugacucagcaacugauacgcgcugcagaaauucgggccucagcaaaccuugccgcgacaaagaugagcgaaugcgugcugggacaauccaagcgggucgacuuuugugguaaaggcuaucaucugaugagcuucccacaguccgcuccacacggcgucguuuuccugcacgugaccuaugugccagcacaggagaagaacuuuacaacagccccggcuaucugccacgauggcaaagcucacuuuccuagagagggaguguuuguaagcaauggaacccauugguucguuacacaaagaaacuuuuaugagccgcaaauuaucacaacagauaauacauucgucuccgggaacugugacguugugauagggauugucaacaacacaguguacgacccccugcaacccgagcuggauucauuuaaagaagaacucgacaaguacuucaagaaucauacuaguccagauguggaucugggcgauauaucaggaaucaaugccagcguggucaauauucaaaaggagauugauagacugaacgagguugccaagaaucugaaugaaagccugaucgaucugcaagaauugggcaaguaugagcaguacauuaaauggcccuggucuggcgggagcggcggaucugggucuggauauauucccgaagcuccuagagauggacaagcuuacguccguaaagacggcgaguggguucuucucuccacauuccucggcugauga (SEQ ID NO: 144)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG**(SEQ ID NO: 145) WuS_3F_2P_pVaxggatccgccaccatggattggacctggattctctttcttgtggcggcggcaacacgcgtccacagcatgttcgtctttctggtattgctgccgcttgtgagtagccagtgtgtgaacttgactaccaggacccagctcccaccggcttataccaattccttcacaagaggtgtctactatccagataaagttttccgcagctcagtgttgcatagcacacaggatctctttctgccattcttcagcaacgtcacgtggtttcatgcgatacacgttagtggaacaaacggaacaaaacgcttcgacaaccctgttctgccattcaatgacggagtttactttgcgagtaccgagaaatctaacatcattagagggtggatctttgggactacattggattctaaaacccagtcactcctcatagtcaataacgctacaaatgtggtgattaaggtatgcgaatttcagttttgcaacgacccatttctcggtgtatattatcacaagaataataaaagttggatggagtccgagttccgcgtctattcatcagccaataattgtactttcgaatatgtttctcaaccgtttctcatggatctcgaaggaaagcaagggaattttaagaatctccgggagtttgtcttcaagaacatagatggctattttaaaatttactcaaagcatactcctattaacctcgttcgggatctcccccagggttttagcgccctggagccactggttgatctgccaattgggattaatatcacacgctttcaaactctcctggcccttcacagatcttacttgaccccaggtgattcaagtagtggttggacagctggagctgctgcatattatgtaggttatctccaaccccgcacctttctcctcaaatataacgagaacggcaccattacagatgcggtggactgtgccttggaccctctttctgagaccaagtgcacactcaaaagcttcactgtcgaaaaggggatttaccagacatcaaattttagagttcaacccaccgaaagcattgtgagatttcctaacattacaaacttgtgcccatttggggaagtctttaacgctacacgctttgctagcgtctatgcttggaaccgaaaacggattagtaactgcgtagctgattattccgtcttgtacaacagcgcatcttttagcactttcaagtgttatggagtaagcccaacaaagctcaacgacctttgttttactaacgtctatgctgattcattcgtgattcgtggggatgaggttcgtcagatcgccccaggccaaaccgggaaaatcgctgattataattataaattgcctgacgattttaccggctgtgtaatcgcctggaattccaacaatcttgattccaaggttggcggcaactacaactatctctaccgtctgtttcgcaaatccaatctcaagcccttcgaacgcgatatttcaactgaaatctatcaggcagggtccactccgtgtaacggcgtagaaggatttaattgttacttcccattgcaaagttatggctttcaacccaccaacggagtcgggtaccaaccatacagagtcgtcgtgctctcatttgagctccttcatgcacctgccacggtgtgcggcccaaagaaatcaacgaaccttgtgaagaataaatgtgtcaattttaactttaatggcctgacagggactggcgtcctcacagaatctaataagaagtttctccctttccagcaatttggtcgcgatatagctgataccacagatgcagttagagacccacagacacttgagattctcgatattaccccgtgctcctttggcggcgtgtccgtcattactcccggtaccaatacgtctaaccaggtagcagtgctctaccaagatgtaaattgtactgaggtacccgtggcaatccatgccgaccaactgactccaacgtggcgggtttattcaaccggaagcaacgtgtttcaaacacgggctggctgccttataggcgctgagcacgtgaataatagttacgagtgtgatatcccgatcggagccggcatctgtgcatcttatcaaacacaaacaaactccccgcgccggcggagaagcgtggctagccaaagtataatcgcttatacaatgtccttgggcgcggaaaattcagtggcttattccaataattcaattgccattcctaccaactttacaattagcgtgaccacagaaatcttgcctgtgtctatgaccaagaccagcgtcgattgcaccatgtatatctgtggagatagcaccgagtgttcaaatttgctcctgcaatacggttccttttgtacacagcttaaccgcgccctcacaggtattgctgttgaacaagacaagaatactcaagaggtatttgctcaggtaaaacaaatttataagaccccaccgataaaagattttggcggtttcaatttctcccaaatattgccagatcctagtaggcgtcgtagatcatttatcgaggatctcctgttcaataaagtaaccctcgccgacgctggtttcatcaaacaatatggcgactgcctgggagatattgcagctagggatttgatttgtgcacagaagttcaatggactcaccgttctcccgcctctcctgacagatgagatgattgcacaatacacctctgctcttttggccgggaccattacgagcggttggacttttggcgcgggtgcggctctccaaattcctttcgcgatgcaaatggcgtatagatttaatggaattggcgttactcaaaacgtcttgtacgagaatcagaaactgatcgccaaccaatttaacagtgcaattggcaaaatccaagatagccttagttctactgcttcagcattgggtaagttgcaagatgtggtcaaccaaaacgcacaagcactcaataccctcgtgaagcaattgtccagcaattttggagctatctcaagtgtgctcaacgacatcctttctaggcttgatccacccgaggcagaggttcaaatcgacagactgataactggcaggctccaatctctgcaaacgtacgtgacacagcaactgattagggctgctgagatcagggcgtccgcgaatttggcagcaaccaaaatgagcgaatgcgtgctgggacaatcaaagagagttgatttctgtggaaagggttaccatctcatgtccttccctcaatcagctccccatggagttgtgtttctgcacgttacttacgtgccggcacaagaaaagaatttcaccactgcaccggctatatgtcatgatgggaaagcccacttcccgcgggaaggcgttttcgtgtccaacgggactcattggttcgtcacacaaaggaacttctatgagccacaaataattacaacagacaacacctttgtctctgggaactgcgatgtcgtgattggaatcgtgaacaacactgtctacgatccgctgcaacccgaactcgactcattcaaagaggaactggataagtatttcaagaaccataccagccccgatgtcgatctgggcgatatctccgggataaatgcttcagtagtaaacattcaaaaggaaatcgaccggctgaacgaggttgcgaagaatcttaatgagtcattgatcgacctgcaagaacttggtaagtatgagcagtacatcaagtggccttggtcaggccgcaggcgtcggcgtcgtgggagcggcggcagtgggagcggatatattccagaagcgccccgagacggacaagcttacgtacgaaaagacggagaatgggtactgctttccacttttcttggctaatga (SEQ ID NO: 146)ggauccgccaccauggauuggaccuggauucucuuucuuguggcggcggcaacacgcguccacagcauguucgucuuucugguauugcugccgcuugugaguagccagugugugaacuugacuaccaggacccagcucccaccggcuuauaccaauuccuucacaagaggugucuacuauccagauaaaguuuuccgcagcucaguguugcauagcacacaggaucucuuucugccauucuucagcaacgucacgugguuucaugcgauacacguuaguggaacaaacggaacaaaacgcuucgacaacccuguucugccauucaaugacggaguuuacuuugcgaguaccgagaaaucuaacaucauuagaggguggaucuuugggacuacauuggauucuaaaacccagucacuccucauagucaauaacgcuacaaauguggugauuaagguaugcgaauuucaguuuugcaacgacccauuucucgguguauauuaucacaagaauaauaaaaguuggauggaguccgaguuccgcgucuauucaucagccaauaauuguacuuucgaauauguuucucaaccguuucucauggaucucgaaggaaagcaagggaauuuuaagaaucuccgggaguuugucuucaagaacauagauggcuauuuuaaaauuuacucaaagcauacuccuauuaaccucguucgggaucucccccaggguuuuagcgcccuggagccacugguugaucugccaauugggauuaauaucacacgcuuucaaacucuccuggcccuucacagaucuuacuugaccccaggugauucaaguagugguuggacagcuggagcugcugcauauuauguagguuaucuccaaccccgcaccuuucuccucaaauauaacgagaacggcaccauuacagaugcgguggacugugccuuggacccucuuucugagaccaagugcacacucaaaagcuucacugucgaaaaggggauuuaccagacaucaaauuuuagaguucaacccaccgaaagcauugugagauuuccuaacauuacaaacuugugcccauuuggggaagucuuuaacgcuacacgcuuugcuagcgucuaugcuuggaaccgaaaacggauuaguaacugcguagcugauuauuccgucuuguacaacagcgcaucuuuuagcacuuucaaguguuauggaguaagcccaacaaagcucaacgaccuuuguuuuacuaacgucuaugcugauucauucgugauucguggggaugagguucgucagaucgccccaggccaaaccgggaaaaucgcugauuauaauuauaaauugccugacgauuuuaccggcuguguaaucgccuggaauuccaacaaucuugauuccaagguuggcggcaacuacaacuaucucuaccgucuguuucgcaaauccaaucucaagcccuucgaacgcgauauuucaacugaaaucuaucaggcaggguccacuccguguaacggcguagaaggauuuaauuguuacuucccauugcaaaguuauggcuuucaacccaccaacggagucggguaccaaccauacagagucgucgugcucucauuugagcuccuucaugcaccugccacggugugcggcccaaagaaaucaacgaaccuugugaagaauaaaugugucaauuuuaacuuuaauggccugacagggacuggcguccucacagaaucuaauaagaaguuucucccuuuccagcaauuuggucgcgauauagcugauaccacagaugcaguuagagacccacagacacuugagauucucgauauuaccccgugcuccuuuggcggcguguccgucauuacucccgguaccaauacgucuaaccagguagcagugcucuaccaagauguaaauuguacugagguacccguggcaauccaugccgaccaacugacuccaacguggcggguuuauucaaccggaagcaacguguuucaaacacgggcuggcugccuuauaggcgcugagcacgugaauaauaguuacgagugugauaucccgaucggagccggcaucugugcaucuuaucaaacacaaacaaacuccccgcgccggcggagaagcguggcuagccaaaguauaaucgcuuauacaauguccuugggcgcggaaaauucaguggcuuauuccaauaauucaauugccauuccuaccaacuuuacaauuagcgugaccacagaaaucuugccugugucuaugaccaagaccagcgucgauugcaccauguauaucuguggagauagcaccgaguguucaaauuugcuccugcaauacgguuccuuuuguacacagcuuaaccgcgcccucacagguauugcuguugaacaagacaagaauacucaagagguauuugcucagguaaaacaaauuuauaagaccccaccgauaaaagauuuuggcgguuucaauuucucccaaauauugccagauccuaguaggcgucguagaucauuuaucgaggaucuccuguucaauaaaguaacccucgccgacgcugguuucaucaaacaauauggcgacugccugggagauauugcagcuagggauuugauuugugcacagaaguucaauggacucaccguucucccgccucuccugacagaugagaugauugcacaauacaccucugcucuuuuggccgggaccauuacgagcgguuggacuuuuggcgcgggugcggcucuccaaauuccuuucgcgaugcaaauggcguauagauuuaauggaauuggcguuacucaaaacgucuuguacgagaaucagaaacugaucgccaaccaauuuaacagugcaauuggcaaaauccaagauagccuuaguucuacugcuucagcauuggguaaguugcaagauguggucaaccaaaacgcacaagcacucaauacccucgugaagcaauuguccagcaauuuuggagcuaucucaagugugcucaacgacauccuuucuaggcuugauccacccgaggcagagguucaaaucgacagacugauaacuggcaggcuccaaucucugcaaacguacgugacacagcaacugauuagggcugcugagaucagggcguccgcgaauuuggcagcaaccaaaaugagcgaaugcgugcugggacaaucaaagagaguugauuucuguggaaaggguuaccaucucauguccuucccucaaucagcuccccauggaguuguguuucugcacguuacuuacgugccggcacaagaaaagaauuucaccacugcaccggcuauaugucaugaugggaaagcccacuucccgcgggaaggcguuuucguguccaacgggacucauugguucgucacacaaaggaacuucuaugagccacaaauaauuacaacagacaacaccuuugucucugggaacugcgaugucgugauuggaaucgugaacaacacugucuacgauccgcugcaacccgaacucgacucauucaaagaggaacuggauaaguauuucaagaaccauaccagccccgaugucgaucugggcgauaucuccgggauaaaugcuucaguaguaaacauucaaaaggaaaucgaccggcugaacgagguugcgaagaaucuuaaugagucauugaucgaccugcaagaacuugguaaguaugagcaguacaucaaguggccuuggucaggccgcaggcgucggcgucgugggagcggcggcagugggagcggauauauuccagaagcgccccgagacggacaagcuuacguacgaaaagacggagaauggguacugcuuuccacuuuucuuggcuaauga (SEQ ID NO: 147)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDPPEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 148) WuS_3F_D2P_GlyDSol_pVaxggatccgccaccatggattggacctggatcttgtttctcgtcgcagctgccacaagagtccacagtatgtttgtatttctcgttctcctgcctcttgtgtcctctcagtgtgtgaatctgacgaccagaactcagctcccacccgcatacactaacagtatcacgaggggcgtttattacccagacaaggttttccgaagcagttgtctgtatagtacacaggatttgttcctgccattctttagtaacgtgacttggtttcatgcgatccacgtttctggcacaaacggtaccaaaagattcgataaccccgtgctgcctttcaatgacggcgtgtatttcgcttctactgaaaagtccggcatcatccggggatggatcttcggcactactttggatagcaaaacccaatccctgctgattgtgaataatgcaaccaacgtggtgatcaaagtctgtgagtttcaattctgtaatgacccatttctgggcgtttactaccacaagaacaacaaaagttggatggagtccgaatttcgcgtctactcatccgcgaacaattgcacatttgagtatgtgagtcaaccattcttgatggatcttgaagggaagcaaggcaatttcaagaacctgcgggagtttgtatttaagaacattgacggctatttcaagatatattctaaacatactcccattaacctcgtgcgcgacttgccccagggattcagtgctctggaaccactggtcgatctccccatcggcattaatattacacgctttcaaactctcctcgctttgcatcggtcctatcttactccgggagatagctcaagcggatggacggcaggcgcggcggcatattatgttggatatctccaaccacgcacgttccttctgaaatataatgaaaatggcactattactgatgcagtcgactgcgccctggaccctctgtctgagaccaagtgtaccctgaagtcattcaccgttgagaagggaatctaccaaacctctaactttagggtacaacctactgagagcattgtccgcttcccgaatatcaccaatctttgtccgtttggtgaggtgtttaacgcgacacgatttgcttccgtatacgcctggaatcgcaaacgcatcagcaactgtgtggctgattactcatttctctataattccgcttccttctctactttcaagtgttatggggtgaatgggactaaacttaatgacttgtgctttactaacgtgtatgctgatagcttcgtcattcgtggagatgaggtcaggcaaatagctcccggacaaacagggaagatagcggactataactacaaactgcctgatgatttcaccgggtgcgtcatcgcgtggaacagcaataacctcgatagcaaggttggcggaaactataattatctctatcgtttgtttaggaaaagcaatctcaagcccttcgagcgggatattaatacaacgatatatcaagctggctctaccccgtgcaacggagtagagggcttcaattgctactttcctttgcagtcctacggattccaacccaccaacggagtgggctaccaaccataccgtgtcgtggttttgagtttcgaactgaaccacgcaccagcaacagtctgcggaccgaagaagagtacaaaccttgtgaagaataagtgcgtgaactttaatttcaatggcctgactggaaccggagttctgacggaatccaataagaaatttctgccgtttcagcaatttggacgggattgtgctggaacgactgatgccgtacgtgatcctcaaacactggaaatcctggacataaccccttgttcctttggtggcgtaagcgttattactccaggcacaaacacatcaaatcaagtcgccgtactgtatcaaggtgtcaactgtactgaagtacctgtagccattcatgcaggacaactgacccctacatggcgagtgtattcaacgggaagcaacgtatttcaaaccagggccggctgtctcatcggagcagagcatgtcaataatagttatgaatgcgacatcccaataggtgctgggatctgcgcgagctaccaaacccaaactaatagcccacgaagacggagatctgtcgcgtcccaaagcattattgcgtacacgatgagcctctgtgcagaaaattcagttgcctacagcaacaatagcatcgctattccaaccaatttcactatcagctgtacaacagaaattctcccagtctccatgacgaagacatcctgcgattgtacaatgtatatatgcggcgactcaacagaatgttcaaatttgttgctgcaatacgggtccttctgcacccaactcaatcgagctcttacaggcatagcggtcgaacaagacaagaacacacaagaagtgtttgcccaagttaaacagtgttacaagacaccacctatcaaagatttcggcggttttaacttttctcagatcttgccagacccatctaggcggcggcgatcctttatcgaggaccttctcttcaataaggtaactcttgcagacgctggatttattaagcaatacggcgactgtctcggggatatcgccgctagggatctgatctgtgcccagaaatttaacggctgcacggtgctgccccctctgctgactgatgaaatgatagcacaatatacttctgcattgctggccggtaccattacatcaggatggacatttggtgccggggcggcgctccaaattcccttcgccatgcaaatggcctataggtttaacggcatcggggtgacccaaaacgtcctctatgagaatcaaaagctgattgctaaccagtttaactcagcaataggaaagattcaagactctctgtcaagtaccgcatccgcccttggaaagctccaagacgttgttaaccagaatgcacaagctctcaacacgctcgtgaaacaactctcttcaaattttggtgcgatctcttctggcccaaatgacattttgagccggcttcccaaggtagaagctgaagtacaaattgatcgcctgatcaccggacggctccaaagtctgcagacgtacgtcacccagcaactgatacgggcagcggagatccgggcttctgccaacctggccgccacgaagatgagcgaatgcgtgctcggacagtccaaaagagtagatttctgtggcaagggctatcatctcatgtcctttccccaatccgcccctcacggagttgtcttccttcattgcacttacgtccccgctcaagaaaagaattttactacggcacctgctatctgtcacgacgggaaagcccattttcctagagaaggtgtgtttgtatctaacggcacgcactggttcgtcacgcaacgtaacttttacgagccccagatcatcaccacagacaatacggatgtatcaggtaattgtgatgtcctgattggtatcgtcaataacactgtatacgatcctttgcaaccggaactggactcctttaaagaggaacttgataagtatttcaagaatcacacttccccagatgtcgatctcggggacatctcaggaattaatgcatcagtggtcaatattcaaaaggaaattgatcgcttgaatgaggttgcaaagaatttgaatgaaagccttatcgaccttcaagagctgggcaaatatgagcagtacattaaatggccttggagcggtcgccggcgccgaaggcggggttccggcggtagcggtagcggttatattccagaagctcctcgcgatgggcaggcttatgtgaggaaagatggtgaatgggtccttttgtccacgttcctcgggtagtaa (SEQ ID NO: 149)ggauccgccaccauggauuggaccuggaucuuguuucucgucgcagcugccacaagaguccacaguauguuuguauuucucguucuccugccucuuguguccucucagugugugaaucugacgaccagaacucagcucccacccgcauacacuaacaguaucacgaggggcguuuauuacccagacaagguuuuccgaagcaguugucuguauaguacacaggauuuguuccugccauucuuuaguaacgugacuugguuucaugcgauccacguuucuggcacaaacgguaccaaaagauucgauaaccccgugcugccuuucaaugacggcguguauuucgcuucuacugaaaaguccggcaucauccggggauggaucuucggcacuacuuuggauagcaaaacccaaucccugcugauugugaauaaugcaaccaacguggugaucaaagucugugaguuucaauucuguaaugacccauuucugggcguuuacuaccacaagaacaacaaaaguuggauggaguccgaauuucgcgucuacucauccgcgaacaauugcacauuugaguaugugagucaaccauucuugauggaucuugaagggaagcaaggcaauuucaagaaccugcgggaguuuguauuuaagaacauugacggcuauuucaagauauauucuaaacauacucccauuaaccucgugcgcgacuugccccagggauucagugcucuggaaccacuggucgaucuccccaucggcauuaauauuacacgcuuucaaacucuccucgcuuugcaucgguccuaucuuacuccgggagauagcucaagcggauggacggcaggcgcggcggcauauuauguuggauaucuccaaccacgcacguuccuucugaaauauaaugaaaauggcacuauuacugaugcagucgacugcgcccuggacccucugucugagaccaaguguacccugaagucauucaccguugagaagggaaucuaccaaaccucuaacuuuaggguacaaccuacugagagcauuguccgcuucccgaauaucaccaaucuuuguccguuuggugagguguuuaacgcgacacgauuugcuuccguauacgccuggaaucgcaaacgcaucagcaacuguguggcugauuacucauuucucuauaauuccgcuuccuucucuacuuucaaguguuauggggugaaugggacuaaacuuaaugacuugugcuuuacuaacguguaugcugauagcuucgucauucguggagaugaggucaggcaaauagcucccggacaaacagggaagauagcggacuauaacuacaaacugccugaugauuucaccgggugcgucaucgcguggaacagcaauaaccucgauagcaagguuggcggaaacuauaauuaucucuaucguuuguuuaggaaaagcaaucucaagcccuucgagcgggauauuaauacaacgauauaucaagcuggcucuaccccgugcaacggaguagagggcuucaauugcuacuuuccuuugcaguccuacggauuccaacccaccaacggagugggcuaccaaccauaccgugucgugguuuugaguuucgaacugaaccacgcaccagcaacagucugcggaccgaagaagaguacaaaccuugugaagaauaagugcgugaacuuuaauuucaauggccugacuggaaccggaguucugacggaauccaauaagaaauuucugccguuucagcaauuuggacgggauugugcuggaacgacugaugccguacgugauccucaaacacuggaaauccuggacauaaccccuuguuccuuugguggcguaagcguuauuacuccaggcacaaacacaucaaaucaagucgccguacuguaucaaggugucaacuguacugaaguaccuguagccauucaugcaggacaacugaccccuacauggcgaguguauucaacgggaagcaacguauuucaaaccagggccggcugucucaucggagcagagcaugucaauaauaguuaugaaugcgacaucccaauaggugcugggaucugcgcgagcuaccaaacccaaacuaauagcccacgaagacggagaucugucgcgucccaaagcauuauugcguacacgaugagccucugugcagaaaauucaguugccuacagcaacaauagcaucgcuauuccaaccaauuucacuaucagcuguacaacagaaauucucccagucuccaugacgaagacauccugcgauuguacaauguauauaugcggcgacucaacagaauguucaaauuuguugcugcaauacggguccuucugcacccaacucaaucgagcucuuacaggcauagcggucgaacaagacaagaacacacaagaaguguuugcccaaguuaaacaguguuacaagacaccaccuaucaaagauuucggcgguuuuaacuuuucucagaucuugccagacccaucuaggcggcggcgauccuuuaucgaggaccuucucuucaauaagguaacucuugcagacgcuggauuuauuaagcaauacggcgacugucucggggauaucgccgcuagggaucugaucugugcccagaaauuuaacggcugcacggugcugcccccucugcugacugaugaaaugauagcacaauauacuucugcauugcuggccgguaccauuacaucaggauggacauuuggugccggggcggcgcuccaaauucccuucgccaugcaaauggccuauagguuuaacggcaucggggugacccaaaacguccucuaugagaaucaaaagcugauugcuaaccaguuuaacucagcaauaggaaagauucaagacucucugucaaguaccgcauccgcccuuggaaagcuccaagacguuguuaaccagaaugcacaagcucucaacacgcucgugaaacaacucucuucaaauuuuggugcgaucucuucuggcccaaaugacauuuugagccggcuucccaagguagaagcugaaguacaaauugaucgccugaucaccggacggcuccaaagucugcagacguacgucacccagcaacugauacgggcagcggagauccgggcuucugccaaccuggccgccacgaagaugagcgaaugcgugcucggacaguccaaaagaguagauuucuguggcaagggcuaucaucucauguccuuuccccaauccgccccucacggaguugucuuccuucauugcacuuacguccccgcucaagaaaagaauuuuacuacggcaccugcuaucugucacgacgggaaagcccauuuuccuagagaagguguguuuguaucuaacggcacgcacugguucgucacgcaacguaacuuuuacgagccccagaucaucaccacagacaauacggauguaucagguaauugugauguccugauugguaucgucaauaacacuguauacgauccuuugcaaccggaacuggacuccuuuaaagaggaacuugauaaguauuucaagaaucacacuuccccagaugucgaucucggggacaucucaggaauuaaugcaucaguggucaauauucaaaaggaaauugaucgcuugaaugagguugcaaagaauuugaaugaaagccuuaucgaccuucaagagcugggcaaauaugagcaguacauuaaauggccuuggagcggucgccggcgccgaaggcgggguuccggcgguagcgguagcgguuauauuccagaagcuccucgcgaugggcaggcuuaugugaggaaagauggugaauggguccuuuuguccacguuccucggguaguaa (SEQ ID NO: 150)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSITRGVYYPDKVFRSSCLYSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSGIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSFLYNSASFSTFKCYGVNGTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDINTTIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELNHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDCAGTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQGVNCTEVPVAIHAGQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLCAENSVAYSNNSIAIPTNFTISCTTEILPVSMTKTSCDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQCYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGCTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSGPNDILSRLPKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHCTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTDVSGNCDVLIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 151) WuS_3F_D2P_GlyD3_pVaxggatccgccaccatggactggacatggatacttttcttggtagcggcggcgacacgcgtgcactccatgtttgttttcctcgtcctgctcccacttgtctcaagtcaatgcgttaacctgactacgaggacgcagctcccgcccgcctacacaaactcttttacccggggtgtgtactaccccgacaaagttttccgcagttcatgtctccactcaacacaggacctctttctgccattcttctcaaatgtcacatggtttcacgccatccacgtttccggcactaacggtaccaaacggttcgacaaccctgttctgccattcaatgatggggtgtattttgcgagcacagagaagtccaatataatcagaggttggatcttcggtacaacgctggacagtaaaactcaatctctgctgatagtgaataacgctacgaacgtcgtcattaaggtgtgcgagtttcaattttgcaacgatccattcttgggagtgtactatcataagaacaacaaatcatggatggagagcgagtttagggtgtattcctctgcaaacaactgtacatttgaatacgtgagccagccttttcttatggacctcgaaggtaagcaaggtaacttcaagaacttgcgggaatttgttttcaagaacattgatggatacttcaaaatttactccaaacatacccctatcaatctggtccgcgaccttccacaaggattttccgcacttgaacccttggtcgacctgcctattggaatcaatatcacgcggtttcagacgcttctcgctctccatagatcctacctcacgcccggcgacagttcaagtgggtggaccgcaggcgcggcggcctattatgtgggatacttgcaaccccgcacttttctcctgaaatataatgagaatgggaccataaccgatgcagttgattgtgccttggaccccctgtccgagaccaaatgcacgctgaagtctttcacagtagagaagggaatttaccaaacttccaacttcagagttcaacccacagaatctatcgttcgctttcccaatattacaaatttgtgtccgtttggagaggtgttcaatgctacaaggtttgcttccgtatatgcctggaatcgtaaacgcatctctaattgcgtagcggactactcagttttgtataacagtgctagcttctccactttcaagtgttacggcgttaatgggaccaagctgaatgacctgtgttttaccaacgtgtatgctgactccttcgtaataagaggggatgaggttaggcaaatcgcccctggccagacagggaaaatcgctgattacaattacaagttgccagatgactttaccgggtgtgtcatcgcttggaactccaataatctggattccaaagttggtgggaactataattacctctatcggctgttcagaaaatccaaccttaagcccttcgaaagagatatcaacactacaatttatcaggctggttcaactccgtgtaatggggtcgagggtttcaactgctacttcccgttgcagagttatgggttccagccgacgaatggggtcgggtaccaaccgtacagagtagtagttctgtcctttgagttgaatcatgccccagcaacagtgtgcggcccaaagaaatcaacaaaccttgttaagaataaatgcgtgaacttcaactttaacgggcttactgggactggggtgctcacagaatccaacaagaaattcttgccattccaacaatttggccgcgattgtgcagatacaaccgacgccgtgagagatccccaaacattggagatacttgatatcactccctgctcttttggtggcgtcagcgtcatcaccccaggaaccaatacaagcaatcaagtggctgtcctttatcaagatgtcaattgtaccgaagtcccagtcgcaatacatgcggatcaactgaccccaacatggagagtttactcaacgggatctaacgtgtttcaaactcgtgctggctgcctgataggagcggagcatgtgaataattcctatgaatgcgacattcccattggggctggaatctgtgcatcctatcaaacacaaactaactctccccgccggcggcggagcgtcgccagccaaagcattattgcatatacgatgtccctgtgcgcagaaaattctgttgcatacagcaataactccatcgctatccctacaaactttaccatcagctgtacaaccgaaatcttgcccgtttctatgactaaaacaagttgtgactgcactatgtacatctgtggcgactcaacagagtgttctaaccttctgcttcaatatggatctttctgtacacaacttaatcgcgctctcaccggtatagctgttgagcaagataagaacactcaggaagttttcgcccaagtcaaacaatgttataaaacaccacccataaaagacttcggcggatttaatttctctcaaatactgccggacccatccaggagacgaagaagcttcatagaagatcttctcttcaacaaggtgaccctggccgatgcggggtttatcaagcaatatggcgactgtctcggcgatattgctgcacgcgatctgatatgtgcacagaaattcaatgggtgtaccgtgctcccacctctgctgacagatgaaatgatcgctcaatataccagtgcgctcttggctggaacaattactagtggttggacttttggggctggagccgcactccaaatcccttttgccatgcaaatggcctatcgctttaatgggataggggtcactcagaatgtcttgtatgaaaaccagaagttgattgctaaccaatttaattcagctatagggaaaattcaagacagcctcagtagtactgccagtgccctgggcaaactgcaagatgtcgtgaaccaaaatgctcaagccctgaataccctcgttaagcaacttagctcaaactttggtgcgatttcctcaggccctaatgacatcctctcaaggctgcctaaagtggaagctgaggtccaaatcgatcgcctgattacgggtcgcctgcaatcactccaaacatatgtcacccagcagttgatcagagcggccgagatacgggcatcagcaaatttggcggccacgaaaatgtcagagtgcgtacttggtcaaagtaaaagagttgatttctgcggaaaaggttaccaccttatgtctttcccccagtccgctccacatggagtggtctttctgcattgtacttatgtgccagcccaagaaaagaattttactaccgcccccgctatttgtcatgatggtaaggcgcacttccccagagaaggagtgtttgtgtccaacgggactcactggtttgtgactcaaaggaacttttatgaacctcaaattatcaccacagataacacatttgtgtccgggaattgcgatgtggttatcggcattgttaataataccgtttacgatcccttgcaacctgagttggatagtttcaaggaagaacttgacaaatactttaagaatcacacttccccggatgtagacctcggggacatttccggaattaatgcgagtgttgtgaatatacagaaagagatagaccgactcaacgaggttgctaagaacctcaacgagagccttatcgatcttcaagaactcggcaaatacgagcaatacattaaatggccttggtccggcagaaggagacggcgaaggggaagtggcggcagcggctctggatacatcccggaagctccacgggatgggcaagcatatgttcgcaaggatggagaatgggtccttcttagcaccttcttgggataatga (SEQ ID NO: 152)ggauccgccaccauggacuggacauggauacuuuucuugguagcggcggcgacacgcgugcacuccauguuuguuuuccucguccugcucccacuugucucaagucaaugcguuaaccugacuacgaggacgcagcucccgcccgccuacacaaacucuuuuacccgggguguguacuaccccgacaaaguuuuccgcaguucaugucuccacucaacacaggaccucuuucugccauucuucucaaaugucacaugguuucacgccauccacguuuccggcacuaacgguaccaaacgguucgacaacccuguucugccauucaaugaugggguguauuuugcgagcacagagaaguccaauauaaucagagguuggaucuucgguacaacgcuggacaguaaaacucaaucucugcugauagugaauaacgcuacgaacgucgucauuaaggugugcgaguuucaauuuugcaacgauccauucuugggaguguacuaucauaagaacaacaaaucauggauggagagcgaguuuaggguguauuccucugcaaacaacuguacauuugaauacgugagccagccuuuucuuauggaccucgaagguaagcaagguaacuucaagaacuugcgggaauuuguuuucaagaacauugauggauacuucaaaauuuacuccaaacauaccccuaucaaucugguccgcgaccuuccacaaggauuuuccgcacuugaacccuuggucgaccugccuauuggaaucaauaucacgcgguuucagacgcuucucgcucuccauagauccuaccucacgcccggcgacaguucaaguggguggaccgcaggcgcggcggccuauuaugugggauacuugcaaccccgcacuuuucuccugaaauauaaugagaaugggaccauaaccgaugcaguugauugugccuuggacccccuguccgagaccaaaugcacgcugaagucuuucacaguagagaagggaauuuaccaaacuuccaacuucagaguucaacccacagaaucuaucguucgcuuucccaauauuacaaauuuguguccguuuggagagguguucaaugcuacaagguuugcuuccguauaugccuggaaucguaaacgcaucucuaauugcguagcggacuacucaguuuuguauaacagugcuagcuucuccacuuucaaguguuacggcguuaaugggaccaagcugaaugaccuguguuuuaccaacguguaugcugacuccuucguaauaagaggggaugagguuaggcaaaucgccccuggccagacagggaaaaucgcugauuacaauuacaaguugccagaugacuuuaccgggugugucaucgcuuggaacuccaauaaucuggauuccaaaguuggugggaacuauaauuaccucuaucggcuguucagaaaauccaaccuuaagcccuucgaaagagauaucaacacuacaauuuaucaggcugguucaacuccguguaauggggucgaggguuucaacugcuacuucccguugcagaguuauggguuccagccgacgaauggggucggguaccaaccguacagaguaguaguucuguccuuugaguugaaucaugccccagcaacagugugcggcccaaagaaaucaacaaaccuuguuaagaauaaaugcgugaacuucaacuuuaacgggcuuacugggacuggggugcucacagaauccaacaagaaauucuugccauuccaacaauuuggccgcgauugugcagauacaaccgacgccgugagagauccccaaacauuggagauacuugauaucacucccugcucuuuugguggcgucagcgucaucaccccaggaaccaauacaagcaaucaaguggcuguccuuuaucaagaugucaauuguaccgaagucccagucgcaauacaugcggaucaacugaccccaacauggagaguuuacucaacgggaucuaacguguuucaaacucgugcuggcugccugauaggagcggagcaugugaauaauuccuaugaaugcgacauucccauuggggcuggaaucugugcauccuaucaaacacaaacuaacucuccccgccggcggcggagcgucgccagccaaagcauuauugcauauacgaugucccugugcgcagaaaauucuguugcauacagcaauaacuccaucgcuaucccuacaaacuuuaccaucagcuguacaaccgaaaucuugcccguuucuaugacuaaaacaaguugugacugcacuauguacaucuguggcgacucaacagaguguucuaaccuucugcuucaauauggaucuuucuguacacaacuuaaucgcgcucucaccgguauagcuguugagcaagauaagaacacucaggaaguuuucgcccaagucaaacaauguuauaaaacaccacccauaaaagacuucggcggauuuaauuucucucaaauacugccggacccauccaggagacgaagaagcuucauagaagaucuucucuucaacaaggugacccuggccgaugcgggguuuaucaagcaauauggcgacugucucggcgauauugcugcacgcgaucugauaugugcacagaaauucaauggguguaccgugcucccaccucugcugacagaugaaaugaucgcucaauauaccagugcgcucuuggcuggaacaauuacuagugguuggacuuuuggggcuggagccgcacuccaaaucccuuuugccaugcaaauggccuaucgcuuuaaugggauaggggucacucagaaugucuuguaugaaaaccagaaguugauugcuaaccaauuuaauucagcuauagggaaaauucaagacagccucaguaguacugccagugcccugggcaaacugcaagaugucgugaaccaaaaugcucaagcccugaauacccucguuaagcaacuuagcucaaacuuuggugcgauuuccucaggcccuaaugacauccucucaaggcugccuaaaguggaagcugagguccaaaucgaucgccugauuacgggucgccugcaaucacuccaaacauaugucacccagcaguugaucagagcggccgagauacgggcaucagcaaauuuggcggccacgaaaaugucagagugcguacuuggucaaaguaaaagaguugauuucugcggaaaagguuaccaccuuaugucuuucccccaguccgcuccacauggaguggucuuucugcauuguacuuaugugccagcccaagaaaagaauuuuacuaccgcccccgcuauuugucaugaugguaaggcgcacuuccccagagaaggaguguuuguguccaacgggacucacugguuugugacucaaaggaacuuuuaugaaccucaaauuaucaccacagauaacacauuuguguccgggaauugcgaugugguuaucggcauuguuaauaauaccguuuacgaucccuugcaaccugaguuggauaguuucaaggaagaacuugacaaauacuuuaagaaucacacuuccccggauguagaccucggggacauuuccggaauuaaugcgaguguugugaauauacagaaagagauagaccgacucaacgagguugcuaagaaccucaacgagagccuuaucgaucuucaagaacucggcaaauacgagcaauacauuaaauggccuugguccggcagaaggagacggcgaaggggaaguggcggcagcggcucuggauacaucccggaagcuccacgggaugggcaagcauauguucgcaaggauggagaauggguccuucuuagcaccuucuugggauaauga (SEQ ID NO: 153)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSCLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVNGTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDINTTIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELNHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDCADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLCAENSVAYSNNSIAIPTNFTISCTTEILPVSMTKTSCDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQCYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGCTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSGPNDILSRLPKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHCTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 154) WuS_3F_D2P_GlyD2_pVaxggatccgccaccatggattggacatggatactgtttctggtcgctgctgccacacgtgtccacagcatgtttgtcttcttggtgctcttgcctcttgtgagctcccaatgtgtgaatctgactacacgtacgcaacttccgcctgcctacaccaactctttcaccagaggcgtgtattatccggataaggtgttcaggagctcctgccttcattcaacacaggatttgtttctgcctttcttttcaaacgttacttggttccatgccatccacgtgtcaggaacaaatggtaccaagagattcgataacccagttctcccttttaatgatggagtctattttgcaagcactgagaaaagtaatattatacgaggttggattttcggaacgacactcgacagtaaaacacaatccctgttgatagtcaacaatgccacgaacgtagttataaaagtttgcgaatttcaattttgcaacgatcctttcctgggtgtgtactatcacaagaacaacaaatcttggatggaaagcgagtttcgagtgtattcttcagcaaacaactgtactttcgaatatgtttctcaaccattcctgatggatctcgaaggtaaacagggcaactttaagaatctgagagagtttgtgtttaagaacattgacggctattttaagatttacagcaaacatacgcctataaaccttgtgagagacctgcctcaagggtttagcgccctggaaccactcgtggacctgcctatcggcatcaatattaccagatttcaaacgctccttgccctgcataggagctatttgacacctggggactcttctagcggctggactgcaggcgctgccgcttattacgtgggatatctccagcctagaactttcctcttgaaatacaacgagaatggaaccataacagacgcagttgattgtgctctcgaccccttgtccgagaccaaatgcacactgaaaagttttaccgtggagaaagggatctatcaaactagtaatttccgcgttcaacccactgagagcatagtgaggtttcctaacattacaaatctttgcccgtttggggaagtgtttaatgccactcgttttgctagtgtatacgcctggaatcgaaagcggatttccaattgcgttgctgactacagtgtactctataatagcgcttcatttagcaccttcaagtgctacggggttaacgggaccaaactcaatgacctctgcttcacgaacgtttacgccgactcctttgtcattcgaggtgacgaagtaagacaaatcgccccaggccagactggaaagatcgcggactacaactataagctgccagacgacttcactggatgtgtgatcgcctggaatagtaacaacctcgactccaaggtgggtggcaattacaattatctctataggctgttcaggaagagtaatttgaaaccattcgagcgcgacataaatacaacaatctaccaagcgggttctaccccttgcaacggcgtggaaggttttaattgttacttccctctccaaagctacgggtttcaaccaacaaacggcgtgggataccaaccatacagggtggttgtgttgagcttcgaattgaatcatgcacctgcaacagtgtgtgggcccaagaagtccaccaatctcgttaagaataaatgcgtgaacttcaactttaacgggttgacagggaccggcgtgcttacggaaagtaataagaaattccttcccttccagcaatttggtcgcgactgtgcggatacaacggacgcagtgcgagacccacagacattggagatcctggacataacaccttgctcttttggcggggtctccgtaataacacctggaacaaataccagcaatcaagtagcggtcttgtatcaagatgtaaactgtactgaagtcccagttgctatacatgcagaccaacttacaccgacgtggcgcgtgtattctacgggctccaacgtattccaaaccagagcagggtgcttgataggggcagagcacgtcaacaatagctatgagtgtgatatcccgataggtgctggaatctgcgcaagttaccagacccaaaccaatagcccccgccggagacgatcagtggcaagccagtctataatagcctacacgatgtcactgtgtgccgaaaatagcgttgcctatagtaacaatagcatcgccattccaaccaatttcacaatatcagtcactactgagattctgcctgtgtcaatgactaaaactagtgtggactgcacaatgtatatttgcggcgattccacagaatgtagcaatcttctgctgcaatatgggagtttctgtacacaattgaatcgggcccttactggaatcgccgtagagcaggacaagaacacccaagaagtctttgcgcaagtcaaacaatgttataagactcccccaattaaagattttggcggctttaattttagccaaatacttcccgaccccagccgccgacgacgctcctttatcgaagatctgttgtttaataaagtcacattggctgatgctggctttatcaaacaatacggtgattgtctgggtgatattgcagcccgagatctgatctgcgcccaaaagtttaacggcttgaccgttctcccgccactcctgacagatgagatgatcgcgcaatatacctctgcactcctggcgggaacaatcactagtggttggactttcggcgccggcgctgcactgcaaattcccttcgccatgcaaatggcctatcggtttaacggaattggtgtgactcagaatgtgctttacgaaaatcagaaactcatagctaatcagtttaacagcgcaatcgggaaaattcaggattccctcagcagcaccgctagcgccttgggcaagctgcaggacgttgtaaaccagaacgctcaggccctcaacactctcgttaaacaattgagctctaactttggggccataagcagtggtcctaacgacatcctgagtcgtctgccaaaggtagaggccgaagtgcaaatcgaccggctcatcactggaagactgcaaagcctgcaaacctatgtcacacagcaacttatacgggccgccgaaatcagggcctcagcaaacctcgcagcaacaaagatgagcgagtgtgtgctgggccaatccaagcgcgtggacttctgtggtaagggataccatctgatgtcctttccccaatccgcgcctcatggagtagttttcctgcacgttacgtatgtgcctgcccaagagaagaactttacaacagcaccagccatttgtcatgacggaaaagcccattttcctagagaaggagtctttgtttccaatgggacacattggtttgttacccagcgtaacttttatgagccacaaatcatcaccacggacaatactttcgtgagcggtaattgtgatgtggtcattggcatagtgaataacactgtttacgaccccctgcaaccggaattggacagcttcaaagaagaactggacaagtacttcaagaaccacacatccccagacgtagacctcggagatatttccggaattaacgcatcagtagttaacatccagaaagaaatagatcgactgaatgaggtcgctaagaacttgaacgaatcacttatagatctccaggaactcggcaaatatgagcaatatattaaatggccctggtcaggtcgcagaagacgccgccggggttccggcggatctggatctggatatattcccgaagctccacgggatgggcaagcctacgtaagaaaggatggagaatgggtacttttgtccacgttcttgggctagtag (SEQ ID NO: 155)ggauccgccaccauggauuggacauggauacuguuucuggucgcugcugccacacguguccacagcauguuugucuucuuggugcucuugccucuugugagcucccaaugugugaaucugacuacacguacgcaacuuccgccugccuacaccaacucuuucaccagaggcguguauuauccggauaagguguucaggagcuccugccuucauucaacacaggauuuguuucugccuuucuuuucaaacguuacuugguuccaugccauccacgugucaggaacaaaugguaccaagagauucgauaacccaguucucccuuuuaaugauggagucuauuuugcaagcacugagaaaaguaauauuauacgagguuggauuuucggaacgacacucgacaguaaaacacaaucccuguugauagucaacaaugccacgaacguaguuauaaaaguuugcgaauuucaauuuugcaacgauccuuuccuggguguguacuaucacaagaacaacaaaucuuggauggaaagcgaguuucgaguguauucuucagcaaacaacuguacuuucgaauauguuucucaaccauuccugauggaucucgaagguaaacagggcaacuuuaagaaucugagagaguuuguguuuaagaacauugacggcuauuuuaagauuuacagcaaacauacgccuauaaaccuugugagagaccugccucaaggguuuagcgcccuggaaccacucguggaccugccuaucggcaucaauauuaccagauuucaaacgcuccuugcccugcauaggagcuauuugacaccuggggacucuucuagcggcuggacugcaggcgcugccgcuuauuacgugggauaucuccagccuagaacuuuccucuugaaauacaacgagaauggaaccauaacagacgcaguugauugugcucucgaccccuuguccgagaccaaaugcacacugaaaaguuuuaccguggagaaagggaucuaucaaacuaguaauuuccgcguucaacccacugagagcauagugagguuuccuaacauuacaaaucuuugcccguuuggggaaguguuuaaugccacucguuuugcuaguguauacgccuggaaucgaaagcggauuuccaauugcguugcugacuacaguguacucuauaauagcgcuucauuuagcaccuucaagugcuacgggguuaacgggaccaaacucaaugaccucugcuucacgaacguuuacgccgacuccuuugucauucgaggugacgaaguaagacaaaucgccccaggccagacuggaaagaucgcggacuacaacuauaagcugccagacgacuucacuggaugugugaucgccuggaauaguaacaaccucgacuccaagguggguggcaauuacaauuaucucuauaggcuguucaggaagaguaauuugaaaccauucgagcgcgacauaaauacaacaaucuaccaagcggguucuaccccuugcaacggcguggaagguuuuaauuguuacuucccucuccaaagcuacggguuucaaccaacaaacggcgugggauaccaaccauacagggugguuguguugagcuucgaauugaaucaugcaccugcaacagugugugggcccaagaaguccaccaaucucguuaagaauaaaugcgugaacuucaacuuuaacggguugacagggaccggcgugcuuacggaaaguaauaagaaauuccuucccuuccagcaauuuggucgcgacugugcggauacaacggacgcagugcgagacccacagacauuggagauccuggacauaacaccuugcucuuuuggcggggucuccguaauaacaccuggaacaaauaccagcaaucaaguagcggucuuguaucaagauguaaacuguacugaagucccaguugcuauacaugcagaccaacuuacaccgacguggcgcguguauucuacgggcuccaacguauuccaaaccagagcagggugcuugauaggggcagagcacgucaacaauagcuaugagugugauaucccgauaggugcuggaaucugcgcaaguuaccagacccaaaccaauagcccccgccggagacgaucaguggcaagccagucuauaauagccuacacgaugucacugugugccgaaaauagcguugccuauaguaacaauagcaucgccauuccaaccaauuucacaauaucagucacuacugagauucugccugugucaaugacuaaaacuaguguggacugcacaauguauauuugcggcgauuccacagaauguagcaaucuucugcugcaauaugggaguuucuguacacaauugaaucgggcccuuacuggaaucgccguagagcaggacaagaacacccaagaagucuuugcgcaagucaaacaauguuauaagacucccccaauuaaagauuuuggcggcuuuaauuuuagccaaauacuucccgaccccagccgccgacgacgcuccuuuaucgaagaucuguuguuuaauaaagucacauuggcugaugcuggcuuuaucaaacaauacggugauugucugggugauauugcagcccgagaucugaucugcgcccaaaaguuuaacggcuugaccguucucccgccacuccugacagaugagaugaucgcgcaauauaccucugcacuccuggcgggaacaaucacuagugguuggacuuucggcgccggcgcugcacugcaaauucccuucgccaugcaaauggccuaucgguuuaacggaauuggugugacucagaaugugcuuuacgaaaaucagaaacucauagcuaaucaguuuaacagcgcaaucgggaaaauucaggauucccucagcagcaccgcuagcgccuugggcaagcugcaggacguuguaaaccagaacgcucaggcccucaacacucucguuaaacaauugagcucuaacuuuggggccauaagcagugguccuaacgacauccugagucgucugccaaagguagaggccgaagugcaaaucgaccggcucaucacuggaagacugcaaagccugcaaaccuaugucacacagcaacuuauacgggccgccgaaaucagggccucagcaaaccucgcagcaacaaagaugagcgagugugugcugggccaauccaagcgcguggacuucugugguaagggauaccaucugauguccuuuccccaauccgcgccucauggaguaguuuuccugcacguuacguaugugccugcccaagagaagaacuuuacaacagcaccagccauuugucaugacggaaaagcccauuuuccuagagaaggagucuuuguuuccaaugggacacauugguuuguuacccagcguaacuuuuaugagccacaaaucaucaccacggacaauacuuucgugagcgguaauugugauguggucauuggcauagugaauaacacuguuuacgacccccugcaaccggaauuggacagcuucaaagaagaacuggacaaguacuucaagaaccacacauccccagacguagaccucggagauauuuccggaauuaacgcaucaguaguuaacauccagaaagaaauagaucgacugaaugaggucgcuaagaacuugaacgaaucacuuauagaucuccaggaacucggcaaauaugagcaauauauuaaauggcccuggucaggucgcagaagacgccgccgggguuccggcggaucuggaucuggauauauucccgaagcuccacgggaugggcaagccuacguaagaaaggauggagaauggguacuuuuguccacguucuugggcuaguag (SEQ ID NO: 156)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSCLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVNGTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDINTTIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELNHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDCADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLCAENSVAYSNNSIAIPTNFTISVTTEILPVSMTKTSVDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQCYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSGPNDILSRLPKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 157) WuS_3F_D2P_GlyD1_pVaxggatccgccaccatggactggacatggatacttttcttggtggcagctgctacacgcgtccactcaatgttcgtctttctggtgctcttgccactggtgagcagccaatgcgttaacctcaccacacgcacgcagcttccacccgcatacactaactcctttacgcgcggcgtgtactatccagataaagtgttccgaagtagcgtcttgcatagcacccaggatctgtttctcccattctttagcaatgtcacatggttccacgctatccacgtgtctgggacgaatggaactaaacgttttgacaatcctgttcttccttttaacgacggcgtatactttgctagtactgagaagtctaacattatccgcggctggattttcgggacaaccctggactccaaaacccagtctctgctgatagtaaacaatgccaccaacgtcgtcattaaagtgtgcgagtttcaattctgcaacgacccctttctgggtgtctattaccacaagaacaataagtcttggatggagtcagaatttcgtgtctattcttctgccaataattgtacatttgagtatgtttctcaaccctttctcatggacctcgaaggcaagcaggggaattttaagaacctgcgggaattcgtctttaagaatatcgacggctatttcaaaatttacagcaaacacacgcctataaacctcgtgcgagatctcccccaaggcttctcagcattggagccattggtcgacttgccaatcggaattaatatcacaaggtttcagactctgctggccctgcatcgctcctatcttacccctggcgattcctcaagtggctggacggccggcgcagcagcctattacgtcggctatctccagccaaggacgtttcttttgaagtataatgaaaatgggactattactgacgccgtcgactgcgctttggaccccctgagcgagacaaagtgcacattgaaaagcttcacggtggagaagggtatttatcaaacttccaactttagggtgcaaccaacagagagcatcgtgaggttccctaatatcactaatctctgtccatttggcgaggtgtttaacgcgaccagatttgcaagcgtatatgcctggaataggaagagaataagcaattgtgttgccgattactctgtcttgtataacagcgcatctttcagcacttttaagtgctatggtgtcaacgggacaaaacttaacgatctttgcttcaccaacgtttacgcagactcttttgtcatacgcggagatgaggtccgacaaatagctcccggccagactgggaaaatcgctgattataactataagcttccagatgacttcacaggatgcgtaattgcatggaactctaacaacctggactcaaaagttggtggcaactataactatctctatcgtttgttccgaaaatcaaaccttaaaccctttgaacgggatattaatacgacaatttaccaagcagggagcactccttgtaacggtgtagaaggtttcaattgttattttcctctgcaatcatacggattccaaccaacaaacggtgtgggttatcaaccttatcgggttgtagttttgagcttcgagcttaaccatgcacccgccacagtatgcggaccgaagaagagtacaaacctggttaagaataaatgtgtaaacttcaactttaatggactgacggggacgggagtactcactgaaagcaataagaaattcttgccttttcagcaattcgggcgggacatagcggacactacagacgccgtgcgcgacccccagactctcgaaatcctggacataaccccgtgctcatttggcggagtttcagtcatcactccagggaccaatacctcaaaccaagtagctgtgctgtatcaagatgtgaattgcaccgaagtaccagtggccattcacgccgatcagctgaccccgacatggcgggtgtactcaaccggttcaaatgtgtttcaaacaagagcaggttgtcttattggcgctgaacacgtgaataactcctatgaatgcgacatcccaattggtgccggaatctgtgcctcttatcaaacacaaactaattcaccaaggcgtaggcgcagcgtcgcctctcaatcaattatagcctacaccatgtcactgggtgccgaaaactccgtcgcgtacagcaacaatagcattgccatccctaccaacttcaccatcagctgtacaactgagatcctgcctgtatccatgacaaagacatcctgcgattgcactatgtacatctgtggagactctactgagtgtagcaacctcttgctccaatacgggagtttctgtacgcaactcaaccgtgccctcaccggcatagccgtagagcaagataagaatacccaggaagtatttgcccaagtaaagcaaatttataagacgccacccattaaagactttggcggtttcaacttcagtcaaatactgccagacccgtctcgcaggagaaggagttttattgaagacctgctctttaacaaggtgactcttgccgatgctggatttattaaacaatatggggattgtctcggagatatcgctgctcgggatcttatctgcgcgcagaaattcaacgggtgtaccgtgctcccacccttgctcactgacgaaatgatcgcgcaatatacctcagcacttctggcgggaactattacatctggttggacattcggcgcaggggcagctctccaaattcccttcgcaatgcaaatggcttacaggttcaatggcataggtgtcacacaaaacgtgctgtacgagaatcaaaagcttatagccaatcagtttaatagcgccataggcaagatccaagattccctgagctccacggcaagcgctctgggaaaattgcaagacgtagtcaatcaaaacgctcaagcgctgaatacccttgtgaaacaactttcttcaaactttggagctatctcatctgggcccaacgatattctgagtcgactgccaaaggttgaagctgaagtccaaattgatcggttgatcacaggaaggctgcaatccctgcagacttacgtgacccagcaactgatcagggcagccgaaataagggcttccgccaatctggcagccacaaagatgtctgaatgtgtcttgggtcaaagcaaacgcgtcgatttctgtggcaaggggtaccatctgatgtcattccctcaatctgcccctcacggtgtggtatttctccattgcacttatgttcccgcacaggagaagaacttcacaacagctcccgccatttgccacgacggaaaggcgcattttccccgcgaaggtgtcttcgtgtccaatgggactcattggtttgtgactcagaggaatttctatgagccgcagattatcaccaccgacaacactttcgtctccggtaactgcgacgtcgttatcggaatcgtcaataacacagtgtatgatcctctgcagccggagctggactcattcaaagaggagttggataaatattttaagaatcatacaagccccgacgtcgatctgggcgatattagtggtatcaatgcgtccgtggttaacattcagaaagagattgacagactcaatgaggtcgccaagaacttgaacgaatccttgattgatctccaggagttgggcaagtatgagcaatatatcaagtggccatggtctgggcgaaggcgccgtcgcagagggtccggcggtagtggttccgggtacataccagaagctccacgagatggtcaagcttatgtaaggaaagacggagagtgggtcctgcttagcacattcttgggttgataa (SEQ ID NO: 158)ggauccgccaccauggacuggacauggauacuuuucuugguggcagcugcuacacgcguccacucaauguucgucuuucuggugcucuugccacuggugagcagccaaugcguuaaccucaccacacgcacgcagcuuccacccgcauacacuaacuccuuuacgcgcggcguguacuauccagauaaaguguuccgaaguagcgucuugcauagcacccaggaucuguuucucccauucuuuagcaaugucacaugguuccacgcuauccacgugucugggacgaauggaacuaaacguuuugacaauccuguucuuccuuuuaacgacggcguauacuuugcuaguacugagaagucuaacauuauccgcggcuggauuuucgggacaacccuggacuccaaaacccagucucugcugauaguaaacaaugccaccaacgucgucauuaaagugugcgaguuucaauucugcaacgaccccuuucugggugucuauuaccacaagaacaauaagucuuggauggagucagaauuucgugucuauucuucugccaauaauuguacauuugaguauguuucucaacccuuucucauggaccucgaaggcaagcaggggaauuuuaagaaccugcgggaauucgucuuuaagaauaucgacggcuauuucaaaauuuacagcaaacacacgccuauaaaccucgugcgagaucucccccaaggcuucucagcauuggagccauuggucgacuugccaaucggaauuaauaucacaagguuucagacucugcuggcccugcaucgcuccuaucuuaccccuggcgauuccucaaguggcuggacggccggcgcagcagccuauuacgucggcuaucuccagccaaggacguuucuuuugaaguauaaugaaaaugggacuauuacugacgccgucgacugcgcuuuggacccccugagcgagacaaagugcacauugaaaagcuucacgguggagaaggguauuuaucaaacuuccaacuuuagggugcaaccaacagagagcaucgugagguucccuaauaucacuaaucucuguccauuuggcgagguguuuaacgcgaccagauuugcaagcguauaugccuggaauaggaagagaauaagcaauuguguugccgauuacucugucuuguauaacagcgcaucuuucagcacuuuuaagugcuauggugucaacgggacaaaacuuaacgaucuuugcuucaccaacguuuacgcagacucuuuugucauacgcggagaugagguccgacaaauagcucccggccagacugggaaaaucgcugauuauaacuauaagcuuccagaugacuucacaggaugcguaauugcauggaacucuaacaaccuggacucaaaaguugguggcaacuauaacuaucucuaucguuuguuccgaaaaucaaaccuuaaacccuuugaacgggauauuaauacgacaauuuaccaagcagggagcacuccuuguaacgguguagaagguuucaauuguuauuuuccucugcaaucauacggauuccaaccaacaaacgguguggguuaucaaccuuaucggguuguaguuuugagcuucgagcuuaaccaugcacccgccacaguaugcggaccgaagaagaguacaaaccugguuaagaauaaauguguaaacuucaacuuuaauggacugacggggacgggaguacucacugaaagcaauaagaaauucuugccuuuucagcaauucgggcgggacauagcggacacuacagacgccgugcgcgacccccagacucucgaaauccuggacauaaccccgugcucauuuggcggaguuucagucaucacuccagggaccaauaccucaaaccaaguagcugugcuguaucaagaugugaauugcaccgaaguaccaguggccauucacgccgaucagcugaccccgacauggcggguguacucaaccgguucaaauguguuucaaacaagagcagguugucuuauuggcgcugaacacgugaauaacuccuaugaaugcgacaucccaauuggugccggaaucugugccucuuaucaaacacaaacuaauucaccaaggcguaggcgcagcgucgccucucaaucaauuauagccuacaccaugucacugggugccgaaaacuccgucgcguacagcaacaauagcauugccaucccuaccaacuucaccaucagcuguacaacugagauccugccuguauccaugacaaagacauccugcgauugcacuauguacaucuguggagacucuacugaguguagcaaccucuugcuccaauacgggaguuucuguacgcaacucaaccgugcccucaccggcauagccguagagcaagauaagaauacccaggaaguauuugcccaaguaaagcaaauuuauaagacgccacccauuaaagacuuuggcgguuucaacuucagucaaauacugccagacccgucucgcaggagaaggaguuuuauugaagaccugcucuuuaacaaggugacucuugccgaugcuggauuuauuaaacaauauggggauugucucggagauaucgcugcucgggaucuuaucugcgcgcagaaauucaacggguguaccgugcucccacccuugcucacugacgaaaugaucgcgcaauauaccucagcacuucuggcgggaacuauuacaucugguuggacauucggcgcaggggcagcucuccaaauucccuucgcaaugcaaauggcuuacagguucaauggcauaggugucacacaaaacgugcuguacgagaaucaaaagcuuauagccaaucaguuuaauagcgccauaggcaagauccaagauucccugagcuccacggcaagcgcucugggaaaauugcaagacguagucaaucaaaacgcucaagcgcugaauacccuugugaaacaacuuucuucaaacuuuggagcuaucucaucugggcccaacgauauucugagucgacugccaaagguugaagcugaaguccaaauugaucgguugaucacaggaaggcugcaaucccugcagacuuacgugacccagcaacugaucagggcagccgaaauaagggcuuccgccaaucuggcagccacaaagaugucugaaugugucuugggucaaagcaaacgcgucgauuucuguggcaagggguaccaucugaugucauucccucaaucugccccucacggugugguauuucuccauugcacuuauguucccgcacaggagaagaacuucacaacagcucccgccauuugccacgacggaaaggcgcauuuuccccgcgaaggugucuucguguccaaugggacucauugguuugugacucagaggaauuucuaugagccgcagauuaucaccaccgacaacacuuucgucuccgguaacugcgacgucguuaucggaaucgucaauaacacaguguaugauccucugcagccggagcuggacucauucaaagaggaguuggauaaauauuuuaagaaucauacaagccccgacgucgaucugggcgauauuagugguaucaaugcguccgugguuaacauucagaaagagauugacagacucaaugaggucgccaagaacuugaacgaauccuugauugaucuccaggaguugggcaaguaugagcaauauaucaaguggccauggucugggcgaaggcgccgucgcagaggguccggcgguagugguuccggguacauaccagaagcuccacgagauggucaagcuuauguaaggaaagacggagaguggguccugcuuagcacauucuuggguugauaa (SEQ ID NO: 159)MDWTWILFLVAAATRVHSMFVFLVLLPLVSSQCVNLTTRTQLPPAYTNSFTRGVYYPDKVFRSSVLHSTQDLFLPFFSNVTWFHAIHVSGTNGTKRFDNPVLPFNDGVYFASTEKSNIIRGWIFGTTLDSKTQSLLIVNNATNVVIKVCEFQFCNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTPGDSSSGWTAGAAAYYVGYLQPRTFLLKYNENGTITDAVDCALDPLSETKCTLKSFTVEKGIYQTSNFRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVNGTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDINTTIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELNHAPATVCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDIADTTDAVRDPQTLEILDITPCSFGGVSVITPGTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRRRSVASQSIIAYTMSLGAENSVAYSNNSIAIPTNFTISCTTEILPVSMTKTSCDCTMYICGDSTECSNLLLQYGSFCTQLNRALTGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSRRRRSFIEDLLFNKVTLADAGFIKQYGDCLGDIAARDLICAQKFNGCTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQKLIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSGPNDILSRLPKVEAEVQIDRLITGRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHCTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWSGRRRRRRGSGGSGSGYIPEAPRDGQAYVRKDGEWVLLSTFLG** (SEQ ID NO: 160)

In some embodiments therefore, the expressible nucleic acid sequencecomprised in the composition of the disclosure comprises at least about70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 68, SEQ ID NO: 69,SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO:77, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 83, SEQ IDNO: 84, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 90, SEQID NO: 92, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 98,SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 104, SEQ IDNO: 105, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 111,SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 117, SEQ IDNO: 119, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 125,SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 131, SEQ IDNO: 132, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 138,SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 144, SEQ IDNO: 146, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 152,SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 158 or SEQ IDNO: 159, or a functional fragment or variant thereof. In someembodiments, the expressible nucleic acid sequence comprises the nucleicacid sequence of SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO:72, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 78, SEQ IDNO: 80, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 86, SEQID NO: 87, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 93,SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO:101, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 107, SEQID NO: 108, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO:114, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, SEQID NO: 122, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO:128, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 134, SEQID NO: 135, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO:141, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 146, SEQ ID NO: 147, SEQID NO: 149, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO:155, SEQ ID NO: 156, SEQ ID NO: 158 or SEQ ID NO: 159, or a functionalfragment or variant thereof. In some embodiments, the expressiblenucleic acid sequence comprised in the composition of the disclosureencodes a polypeptide comprising at least about 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%sequence identity to SEQ ID NO: 70, SEQ ID NO: 73, SEQ ID NO: 76, SEQ IDNO: 79, SEQ ID NO: 82, SEQ ID NO: 85, SEQ ID NO: 88, SEQ ID NO: 91, SEQID NO: 94, SEQ ID NO: 97, SEQ ID NO: 100, SEQ ID NO: 103, SEQ ID NO:106, SEQ ID NO: 109, SEQ ID NO: 112, SEQ ID NO: 115, SEQ ID NO: 118, SEQID NO: 121, SEQ ID NO: 124, SEQ ID NO: 127, SEQ ID NO: 130, SEQ ID NO:133, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 142, SEQ ID NO: 145, SEQID NO: 148, SEQ ID NO: 151, SEQ ID NO: 154, SEQ ID NO: 157 or SEQ ID NO:160, or a functional fragment or variant thereof. In some embodiments,the expressible nucleic acid sequence encodes a polypeptide comprisingthe amino acid sequence of SEQ ID NO: 70, SEQ ID NO: 73, SEQ ID NO: 76,SEQ ID NO: 79, SEQ ID NO: 82, SEQ ID NO: 85, SEQ ID NO: 88, SEQ ID NO:91, SEQ ID NO: 94, SEQ ID NO: 97, SEQ ID NO: 100, SEQ ID NO: 103, SEQ IDNO: 106, SEQ ID NO: 109, SEQ ID NO: 112, SEQ ID NO: 115, SEQ ID NO: 118,SEQ ID NO: 121, SEQ ID NO: 124, SEQ ID NO: 127, SEQ ID NO: 130, SEQ IDNO: 133, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 142, SEQ ID NO: 145,SEQ ID NO: 148, SEQ ID NO: 151, SEQ ID NO: 154, SEQ ID NO: 157 or SEQ IDNO: 160, or a functional fragment or variant thereof.

B. Nucleic Acid Molecule

In one aspect, the present disclosure also relates to a nucleic acidmolecule that comprises any of the disclosed expressible nucleic acidsequences. In some embodiments, the nucleic acid molecule comprising oneor a plurality of the disclosed expressible nucleic acid sequences is inform of a messanger RNA. In some embodiments, the nucleic acid moleculecomprising one or a plurality of the disclosed expressible nucleic acidsequences is in form of a DNA. In some embodiments, the expressiblenucleic acid sequence disclosed herein can be part of a plasmid and thusthe nucleic acid molecule is a plasmid comprising such an expressiblenucleic acid sequence. In some embodiments, provided herein is a vectoror plasmid that is capable of expressing at least a monomer of aself-assembling nanoparticle and a viral antigen construct or constructsin the cell of a mammal in a quantity effective to elicit an immuneresponse in the mammal. The vector or plasmid may comprise heterologousnucleic acid encoding the one or more viral antigens (such as SARS-CoV-2antigens). In some embodiments, provided herein is a vector or plasmidthat is capable of expressing at least one soluble trimer of acoronavirus or SARS-CoV-2 envelope polypeptide or constructs in the cellof a mammal in a quantity effective to elicit an immune response in themammal. In some embodiments, the nucleic acid expresses a trimer of thespike protein of SARS-CoV-2 or a functional fragment or variant thereof.The vector may be a plasmid. The plasmid may be useful for transfectingcells with nucleic acid encoding a viral antigen, which the transformedhost cell is cultured and maintained under conditions wherein expressionof the viral antigen takes place and wherein the structure of thenanoparticle with the antigen or trimer elicits an immune response of amagnitude greater than and/or more therapeutically effective than theimmune repsonse elicited by the antigen alone. The plasmid may furthercomprise an initiation codon, which may be upstream of the expressiblesequence, and a stop codon, which may be downstream of the codingsequence. The initiation and termination codon may be in frame with theexpressible sequence.

The plasmid may also comprise a promoter that is operably linked to thecoding sequence. The promoter operably linked to the coding sequence maybe a promoter from simian virus 40 (SV40), a mouse mammary tumor virus(MMTV) promoter, a human immunodeficiency virus (HIV) promoter such asthe bovine immunodeficiency virus (BIV) long terminal repeat (LTR)promoter, a Moloney virus promoter, an avian leukosis virus (ALV)promoter, a cytomegalovirus (CMV) promoter such as the CMV immediateearly promoter, Epstein Barr virus (EBV) promoter, or a Rous sarcomavirus (RSV) promoter. The promoter may also be a promoter from a humangene such as human actin, human myosin, human hemoglobin, human musclecreatine, or human metalothionein. The promoter may also be a tissuespecific promoter, such as a muscle or skin specific promoter, naturalor synthetic. Examples of such promoters are described in US patentapplication publication No. US20040175727, the contents of which areincorporated herein in its entirety. The plasmid may also comprise apolyadenylation signal, which may be downstream of the coding sequence.The polyadenylation signal may be a SV40 polyadenylation signal, LTRpolyadenylation signal, bovine growth hormone (bGH) polyadenylationsignal, human growth hormone (hGH) polyadenylation signal, or humanβ-globin polyadenylation signal. The SV40 polyadenylation signal may bea polyadenylation signal from a pCEP4 plasmid (Invitrogen, San Diego,Calif.).

The plasmid may also comprise an enhancer upstream of the codingsequence. The enhancer may be human actin, human myosin, humanhemoglobin, human muscle creatine or a viral enhancer such as one fromCMV, FMDV, RSV or EBV. Polynucleotide function enhancers are describedin U.S. Pat. Nos. 5,593,972, 5,962,428, and WO94/016737, the contents ofeach are fully incorporated by reference. The plasmid may also comprisea mammalian origin of replication in order to maintain the plasmidextrachromosomally and produce multiple copies of the plasmid in a cell.The plasmid may be pVAX1, pCEP4 or pREP4 from ThermoFisher Scientific(San Diego, Calif.), which may comprise the Epstein Barr virus origin ofreplication and nuclear antigen EBNA-1 coding region, which may producehigh copy episomal replication without integration.

In some embodiments, the vector can be pVAX1 or a pVax1 variant withchanges such as the variant plasmid described herein. The variant pVax1plasmid is a 2998 basepair variant of the backbone vector plasmid pVAX1(Invitrogen, Carlsbad Calif.). The CMV promoter is located at bases137-724. The T7 promoter/priming site is at bases 664-683. Multiplecloning sites are at bases 696-811. Bovine GH polyadenylation signal isat bases 829-1053. The Kanamycin resistance gene is at bases 1226-2020.The pUC origin is at bases 2320-2993. The vaccine may comprise theconsensus antigens and plasmids at quantities of from about 1 nanogramto 100 milligrams; about 1 microgram to about 10 milligrams; orpreferably about 0.1 microgram to about 10 milligrams; or morepreferably about 1 milligram to about 2 milligram. In some embodiments,pharmaceutical compositions according to the present disclosure comprisefrom about 1 nanogram to about 1000 micrograms of DNA. The nucleic acidsequence for the pVAX1 backbone sequence is as follows:

(SEQ ID NO: 161)  gactcttcgcgatgtacgggccagatatacgcgttgacattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggactatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctggctaactagagaacccactgcttactggcttatcgaaattaatacgactcactatagggagacccaagctggctagcgtttaaacttaagcttggtaccgagctcggatccactagtccagtgtggtggaattctgcagatatccagcacagtggcggccgctcgagtctagagggcccgtttaaacccgctgatcagcctcgactgtgccttctagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattctggggggtggggtggggcaggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcggtgggctctatggcttctactgggcggttttatggacagcaagcgaaccggaattgccagctggggcgccctctggtaaggttgggaagccctgcaaagtaaactggatggctttctcgccgccaaggatctgatggcgcaggggatcaagctctgatcaagagacaggatgaggatcgtttcgcatgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtccggtgccctgaatgaactgcaagacgaggcagcgcggctatcgtggctggccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcgggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtcatctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgcggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcgaaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcgatcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgttcgccaggctcaaggcgagcatgcccgacggcgaggatctcgtcgtgacccatggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttctggattcatcgactgtggccggctgggtgtggcggaccgctatcaggacatagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggctgaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcatcgccttctatcgccttcttgacgagttcttctgaattattaacgcttacaatttcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatacaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatagcacgtgctaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctgggcttttgctggccttttgctcacatgttctt

Other vectors or plasmids that can be used herein to produce the vaccineof the present disclosure include, but not limited to, pcDNA3.1(+), pCImammalian expression vector, pSI vector, pZeoSV2(+), phCMV1, pTCP andpIRES with their respective backbone sequence as follows.

The pcDNA3.1(+) backbone sequence (SEQ ID NO: 162):

gacggatcgggagatctcccgatcccctatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagtatctgctccctgcttgtgtgttggaggtcgctgagtagtgcgcgagcaaaatttaagctacaacaaggcaaggcttgaccgacaattgcatgaagaatctgcttagggttaggcgttttgcgctgcttcgcgatgtacgggccagatatacgcgttgacattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctctctggctaactagagaacccactgcttactggcttatcgaaattaatacgactcactatagggagacccaagctggctagcgtttaaacttaagcttggtaccgagctcggatccactagtccagtgtggtggaattctgcagatatccagcacagtggcggccgctcgagtctagagggcccgtttaaacccgctgatcagcctcgactgtgccttctagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattctggggggtggggtggggcaggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcggtgggctctatggcttctgaggcggaaagaaccagctggggctctagggggtatccccacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattaattctgtggaatgtgtgtcagttagggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctctgcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagctcccgggagcttgtatatccattttcggatctgatcaagagacaggatgaggatcgtttcgcatgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtccggtgccctgaatgaactgcaggacgaggcagcgcggctatcgtggctggccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcgggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtcatctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgcggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcgaaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcgatcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgttcgccaggctcaaggcgcgcatgcccgacggcgaggatctcgtcgtgacccatggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttctggattcatcgactgtggccggctgggtgtggcggaccgctatcaggacatagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggctgaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcatcgccttctatcgccttcttgacgagttcttctgagcgggactctggggttcgaaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgattccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctggagttcttcgcccaccccaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctgtataccgtcgacctctagctagagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcac atttccccgaaaagtgccacctgacgtc 

The pCI mammalian expression vector backbone sequence (SEQ ID NO: 163):

tcaatattggccattagccatattattcattggttatatagcataaatcaatattggctattggccattgcatacgttgtatctatatcataatatgtacatttatattggctcatgtccaatatgaccgccatgttggcattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtccgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttacgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacaccaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaataaccccgccccgttgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcactagaagctttattgcggtagtttatcacagttaaattgctaacgcagtcagtgcttctgacacaacagtctcgaacttaagctgcagaagttggtcgtgaggcactgggcaggtaagtatcaaggttacaagacaggtttaaggagaccaatagaaactgggcttgtcgagacagagaagactcttgcgtttctgataggcacctattggtcttactgacatccactttgcctttctctccacaggtgtccactcccagttcaattacagctcttaaggctagagtacttaatacgactcactataggctagcctcgagaattcacgcgtggtacctctagagtcgacccgggcggccgcttcgagcagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggagatgtgggaggttttttaaagcaagtaaaacctctacaaatgtggtaaaatcgataaggatccgggctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgcttacaatttcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagccccgacacccgccaacacccgctgacgcgccctgacgggcttgtctgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcatgtgtcagaggttttcaccgtcatcaccgaaacgcgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggcaacaacgttgcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatggctcgac agatct

The pSI vector backbone sequence (SEQ ID NO: 164):

gcgcagcaccatggcctgaaataacctctgaaagaggaacttggttaggtaccttctgaggcggaaagaaccagctgtggaatgtgtgtcagttagggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctcggcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagcttgattcttctgacacaacagtctcgaacttaagctgcagaagttggtcgtgaggcactgggcaggtaagtatcaaggttacaagacaggtttaaggagaccaatagaaactgggcttgtcgagacagagaagactcttgcgtttctgataggcacctattggtcttactgacatccactttgcctttctctccacaggtgtccactcccagttcaattacagctcttaaggctagagtacttaatacgactcactataggctagcctcgagaattcacgcgtggtacctctagagtcgacccgggcggccgcttcgagcagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggaggtgtgggaggttttttaaagcaagtaaaacctctacaaatgtggtaaaatcgataaggatccgggctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgcttacaatttcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagccccgacacccgccaacacccgctgacgcgccctgacgggcttgtctgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcatgtgtcagaggttttcaccgtcatcaccgaaacgcgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggcaacaacgttgcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatggctcgacagatct

The pZeoSV2(+) backbone sequence (SEQ ID NO: 165):

ggatcgatccggctgtggaatgtgtgtcagttagggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctcggcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagctctctggctaactagagaacccactgcttactggcttatcgaaattaatacgactcactatagggagacccaagctggctagcgtttaaacttaagcttggtaccgagctcggatccactagtccagtgtggtggaattctgcagatatccagcacagtggcggccgctcgagtctagagggcccgtttaaacccgctgatcagcctcgactgtgccttctagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattctggggggtggggtggggcaggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcggtgggctctatggcttctgaggcggaaagaaccagcatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgacattaacctataaaaataggcgtatcacgaggccctttcgtctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggctggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgcttacaatttccattcgccattcaggctgaactagatctagagtccgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccgggaccgatccagcctccgcggccgggaacggtgcattggaacggaccgtgttgacaattaatcatcggcatagtatatcggcatagtataatacgacaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgctcaccgcgcgcgacgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggaggacgacttcgccggtgtggtccgggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaacaccctggcctgggtgtgggtgcgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacgaacttccgggacgcctccgggccggccatgaccgagatcggcgagcagccgtgggggcgggagttcgccctgcgcgacccggccggcaactgcgtgcacttcgtggccgaggagcaggactgacactcgacctcgaaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtct

The phCMV1 backbone sequence (SEQ ID NO: 166):

tagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcgcctggagacgccatccacgctgttttgacctccatagaagacaccgggaccgatccagcctccgcggccgggaacggtgcattggaacgcggattccccgtgccaagagtgacgtaagtaccgcctatagactctataggcacacccctttggctcttatgcatgaattaatacgactcactatagggagacagactgttcctttcctgggtcttttctgcaggcaccgtcgtcgacttaacagatctcgagctcaagcttcgaattctgcagtcgacggtaccgcgggcccgggatccaccgggtacaagtaaagcggccgcgactctagatcataatcagccataccacatttgtagaggttttacttgctttaaaaaacctcccacacctccccctgaacctgaaacataaaatgaatgcaattgttgttgttaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttaaggcgtaaattgtaagcgttaatattttgttaaaattcgcgttaaatttttgttaaatcagctcattttttaaccaataggccgaaatcggcaaaatcccttataaatcaaaagaatagaccgagatagggttgagtgttgttccagtttggaacaagagtccactattaaagaacgtggactccaacgtcaaagggcgaaaaaccgtctatcagggcgatggcccactacgtgaaccatcaccctaatcaagttttttggggtcgaggtgccgtaaagcactaaatcggaaccctaaagggagcccccgatttagagcttgacggggaaagccggcgaacgtggcgagaaaggaagggaagaaagcgaaaggagcgggcgctagggcgctggcaagtgtagcggtcacgctgcgcgtaaccaccacacccgccgcgcttaatgcgccgctacagggcgcgtcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaaggaagagtcctgaggcggaaagaaccagctgtggaatgtgtgtcagttagggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctcggcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaagatcgatcaagagacaggatgaggatcgtttcgcatgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtccggtgccctgaatgaactgcaagacgaggcagcgcggctatcgtggctggccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcgggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtcatctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgcggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcgaaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcgatcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgttcgccaggctcaaggcgagcatgcccgacggcgaggatctcgtcgtgacccatggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttctggattcatcgactgtggccggctgggtgtggcggaccgctatcaggacatagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggctgaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcatcgccttctatcgccttcttgacgagttcttctgagcgggactctggggttcgaaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgattccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctggagttcttcgcccaccctagggggaggctaactgaaacacggaaggagacaataccggaaggaacccgcgctatgacggcaataaaaagacagaataaaacgcacggtgttgggtcgtttgttcataaacgcggggttcggtcccagggctggcactctgtcgataccccaccgagaccccattggggccaatacgcccgcgtttcttccttttccccaccccaccccccaagttcgggtgaaggcccagggctcgcagccaacgtcggggcggcaggccctgccatagcctcaggttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgattctgtggataaccgtattaccgccatgcat

The pTCP backbone sequence (SEQ ID NO: 167):

tagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctggtttagtgaaccgtggatcccgtcgcttaccgattcagaatggttgatatccgccattctgaatcggtaagcgacgaagcttaataaaggatcttttattttcattggatctgtgtgttggttttttgtgtgcggccgccctcgactgtgccttctagaagacaatagcaggcatgctggggatgcggtgggctctatggcttctgaggcggaaagaaccagctggggctctagggggtatccccacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattaattctgtggaatgtgtgtcagttagggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctctgcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagctcccgggatgaccgagtacaagcccacggtgcgcctcgccacccgcgacgacgtcccgcgggccgtacgcaccctcgccgccgcgttcgccgactaccccgccacgcgccacaccgtcgacccggaccgccacatcgagcgggtcaccgagctgcaagaactcttcctcacgcgcgtcgggctcgacatcggcaaggtgtgggtcgcggacgacggcgccgcggtggcggtctggaccacgccggagagcgtcgaagcgggggcggtgttcgccgagatcggcccgcgcatggccgagttgagcggttcccggctggccgcgcagcaacagatggaaggcctcctggcgccgcaccggcccaaggagcccgcgtggttcctggccaccgtcggcgtctcgcccgaccaccagggcaagggtctgggcagcgccgtcgtgctccccggagtggaggcggccgagcgcgccggggtgcccgccttcctggagacctccgcgccccgcaacctccccttctacgagcggctcggcttcaccgtcaccgccgacgtcgaggtgcccgaaggaccgcgcacctggtgcatgacccgcaagcccggtgcctgattcgaatgaccgaccaagcgacgcccaacctgccatcacgagatttcgattccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggacgccggctggatgatcctccagcgcggggatctcatgctggagttcttcgcccaccccaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctgtataccgtcgacctctagctagagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtcgacggatcgggagatctcccgatcccctatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagtatctgctccctgcttgtgtgttggaggtcgctgagtagtgcgcgagcaaaatttaagctacaacaaggcaaggcttgaccgacaattgcatgaagaatctgcttagggttaggcgttttgcgctgcttcgcgatgtacgggccagatatacgcgttgacatt gattattgac

The pIRES backbone sequence (SEQ ID NO: 168):

tcaatattggccattagccatattattcattggttatatagcataaatcaatattggctattggccattgcatacgttgtatctatatcataatatgtacatttatattggctcatgtccaatatgaccgccatgttggcattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtccgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttacgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacaccaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactgcgatcgcccgccccgttgacgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagagctcgtttagtgaaccgtcagatcactagaagctttattgcggtagtttatcacagttaaattgctaacgcagtcagtgcttctgacacaacagtctcgaacttaagctgcagtgactctcttaaggtagccttgcagaagttggtcgtgaggcactgggcaggtaagtatcaaggttacaagacaggtttaaggagaccaatagaaactgggcttgtcgagacagagaagactcttgcgtttctgataggcacctattggtcttactgacatccactttgcctttctctccacaggtgtccactcccagttcaattacagctcttaaggctagagtacttaatacgactcactataggctagcctcgagaattcacgcgtcgagcatgcatctagggcggccaattccgcccctctcccccccccccctctccctcccccccccctaacgttactggccgaagccgcttggaataaggccggtgtgcgtttgtctatatgttattttccaccatattgccgtcttttggcaatgtgagggcccggaaacctggccctgtcttcttgacgagcattcctaggggtctttcccctctcgccaaaggaatgcaaggtctgttgaatgtcgtgaaggaagcagttcctctggaagcttcttgaagacaaacaacgtctgtagcgaccctttgcaggcagcggaaccccccacctggcgacaggtgcctctgcggccaaaagccacgtgtataagatacacctgcaaaggcggcacaaccccagtgccacgttgtgagttggatagttgtggaaagagtcaaatggctctcctcaagcgtattcaacaaggggctgaaggatgcccagaaggtaccccattgtatgggatctgatctggggcctcggtgcacatgctttacatgtgtttagtcgaggttaaaaaaacgtctaggccccccgaaccacggggacgtggttttcctttgaaaaacacgatgataagcttgccacaacccgggatcctctagagtcgacccgggcggccgcttccctttagtgagggttaatgcttcgagcagacatgataagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggagatgtgggaggttttttaaagcaagtaaaacctctacaaatgtggtaaaatccgataaggatcgatccgggctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatggcgaatggacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgcttacaatttcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatacgcggatctgcgcagcaccatggcctgaaataacctctgaaagaggaacttggttaggtaccttctgaggcggaaagaaccagctgtggaatgtgtgtcagttagggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcatctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctcggcctctgagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagcttgattcttctgacacaacagtctcgaacttaaggctagagccaccatgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctattcggctatgactgggcacaacagacaatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtccggtgccctgaatgaactgcaggacgaggcagcgcggctatcgtggctggccacgacgggcgttccttgcgcagctgtgctcgacgttgtcactgaagcgggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtcatctcaccttgctcctgccgagaaagtatccatcatggctgatgcaatgcggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcgaaacatcgcatcgagcgagcacgtactcggatggaagccggtcttgtcgatcaggatgatctggacgaagagcatcaggggctcgcgccagccgaactgttcgccaggctcaaggcgcgcatgcccgacggcgaggatctcgtcgtgacccatggcgatgcctgcttgccgaatatcatggtggaaaatggccgcttttctggattcatcgactgtggccggctgggtgtggcggaccgctatcaggacatagcgttggctacccgtgatattgctgaagagcttggcggcgaatgggctgaccgcttcctcgtgctttacggtatcgccgctcccgattcgcagcgcatcgccttctatcgccttcttgacgagttcttctgagcgggactctggggttcgaaatgaccgaccaagcgacgcccaacctgccatcacgatggccgcaataaaatatctttattttcattacatctgtgtgttggttttttgtgtgaatcgatagcgataaggatccgcgtatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagccccgacacccgccaacacccgctgacgcgccctgacgggcttgtctgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcatgtgtcagaggttttcaccgtcatcaccgaaacgcgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggcaacaacgttgcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggcct tttgctcacatggctcgacagatct

In some embodiments therefore, the composition of the disclosurecomprises a nucleic acid molecule comprising at least about 70%, 75%,80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% sequence identity to SEQ ID NO: 161, SEQ ID NO: 162, SEQ IDNO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 1676or SEQ ID NO: 168, or a functional fragment or variant thereof. In someembodiments, the composition of the disclosure comprises a nucleic acidmolecule comprising the nucleotide sequence of SEQ ID NO: 161, SEQ IDNO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166,SEQ ID NO: 1676 or SEQ ID NO: 168, or a functional fragment or variantthereof. In some embodiments, the composition of the disclosurecomprises a nucleic acid molecule that is a pVax variant.

In some embodiments, the composition of the disclosure comprises anucleic acid molecule or a plasmid comprising at least about 70%, 75%,80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%. 99% or 100% sequence identity to SEQ ID NO: 161, SEQ ID NO: 162,SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ IDNO: 1676 or SEQ ID NO: 168, or a functional fragment or variant thereof,and an expressible nucleic acid sequence comprising a first nucleic acidsequence encoding a scaffold domain comprising any of theself-assembling polypeptides disclosed herein, or a functional fragmentor variant thereof, and a second nucleic acid sequence encoding anantigen domain comprising any of the viral antigens disclosed herein, ora functional fragment or variant thereof. In some embodiments, thecomposition of the disclosure comprises a nucleic acid molecule or aplasmid comprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO:164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 1676 or SEQ ID NO: 168,or a functional fragment or variant thereof, and an expressible nucleicacid sequence comprising a first nucleic acid sequence encoding ascaffold domain comprising a self-assembling polypeptide comprising atleast about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 8,SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO:18 or SEQ ID NO: 20, or a functional fragment or variant thereof, and asecond nucleic acid sequence encoding an antigen domain comprising aviral antigen comprising at least about 70%, 75%, 80%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ IDNO: 66, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174,SEQ ID NO: 175, SEQ ID NO: 176 or SEQ ID NO: 177, or a functionalfragment or variant thereof. In some embodiments, the composition of thedisclosure comprises a nucleic acid molecule or a plasmid comprising atleast about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO:161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQID NO: 166, SEQ ID NO: 1676 or SEQ ID NO: 168, or a functional fragmentor variant thereof, and an expressible nucleic acid sequence comprisinga first nucleic acid sequence comprising at least about 70%, 75%, 80%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99% or 100% sequence identity to SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO:11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17 or SEQ ID NO: 19, or afunctional fragment or variant thereof, and a second nucleic acidsequence comprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63 or SEQ ID NO:65, or a functional fragment or variant thereof. In some embodiments,such nucleic acid molecules or plasmids may further comprise a thirdnucleic acid sequence encoding a leader sequence comprising at leastabout 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 1 or SEQID NO: 5, or a functional fragment or variant thereof. In suchembodiments, the third nucleic acid sequence encoding a leader sequencemay comprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 6, ora functional fragment or variant thereof.

In some embodiments, the nucleic acid molecules or plasmids of thedisclosure may additionally comprise another nucleic acid sequenceencoding a linker comprising at least about 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ IDNO: 25, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46,SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO:56 or SEQ ID NO: 58, or a functional fragment or variant thereof. Insome embodiments, the nucleic acid sequence encoding a linker maycomprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 21, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO:30, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ IDNO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55 or SEQ ID NO: 57or a functional fragment or variant thereof.

In some embodiments, the composition of the disclosure comprises anucleic acid molecule or a plasmid comprising at least about 70%, 75%,80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or 100% sequence identity to SEQ ID NO: 161, SEQ ID NO: 162,SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ IDNO: 1676 or SEQ ID NO: 168, or a functional fragment or variant thereof,and an expressible nucleic acid sequence comprising a first nucleic acidsequence encoding a leader sequence comprising any of the leadersequences disclosed herein, or a functional fragment or variant thereof,and a second nucleic acid sequence encoding a viral trimer (or threeviral monomers) comprising any of the viral antigens disclosed herein,or a functional fragment or variant thereof. In some embodiments, thecomposition of the disclosure comprises a nucleic acid molecule or aplasmid comprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO:164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 1676 or SEQ ID NO: 168,or a functional fragment or variant thereof, and an expressible nucleicacid sequence comprising a first nucleic acid sequence encoding a leadersequence comprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 1 or SEQ ID NO: 5, or a functional fragment orvariant thereof, and a second nucleic acid sequence encoding three viralmonomers, each viral monomer independently comprising at least about70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60, SEQ IDNO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO: 172,SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 or SEQ IDNO: 177, or a functional fragment or variant thereof. In someembodiments, the composition of the disclosure comprises a nucleic acidmolecule or a plasmid comprising at least about 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 1676 or SEQ IDNO: 168, or a functional fragment or variant thereof, and an expressiblenucleic acid sequence comprising a first nucleic acid sequencecomprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 6, or afunctional fragment or variant thereof, and a second nucleic acidsequence encoding three viral monomers, each viral monomer independentlybeing encoded by a nucleic acid sequence comprising at least about 70%,75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 59, SEQ ID NO: 61,SEQ ID NO: 63 or SEQ ID NO: 65, or a functional fragment or variantthereof. In some embodiments, each of the viral monomers is linked byone or more linker peptides comprising at least about 70%, 75%, 80%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99% or 100% sequence identity to SEQ ID NO: 22, SEQ ID NO: 23, SEQ IDNO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO: 34, SEQID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44,SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO:54, SEQ ID NO: 56 or SEQ ID NO: 58, or a functional fragment or variantthereof. In some embodiments, each of the viral monomers is linked byone or more linker peptides encoded by a nucleic acid sequencecomprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 21, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO:30, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ IDNO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55 or SEQ ID NO: 57or a functional fragment or variant thereof.

In some embodiments, any of the nucleic acid molecules or plasmids ofthe disclosure additionally comprises a nucleic acid sequence encoding afurin cleavage site comprising at least about 60%, 65%, 70%, 75%, 80%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99% or 100% sequence identity to SEQ ID NO: 67, or a functional fragmentor variant thereof.

In some embodiments, the nucleic acid molecule or plasmid may furthercomprises a nucleic acid encoding a transmembrane domain and a foldondomain. A non-limiting example of the transmembrane domain is thetransmembrane domain of a platelet derived growth factor receptorcomprising the sequence of AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQKKPR (SEQ ID NO: 169). A non-limitingexample of the foldon domain may comprise the sequence ofYIPEAPRDGQAYVRKD GEWVLLSTFL (SEQ ID NO: 170). Thus, in some embodiments,the composition of the disclosure comprises a nucleic acid molecule or aplasmid comprising a nucleic acid sequence encoding a transmembranedomain comprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 169, or a functional fragment or variant thereof.In some embodiments, the composition of the disclosure comprises anucleic acid molecule or a plasmid comprising a nucleic acid sequenceencoding a foldon domain comprising at least about 70%, 75%, 80%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or100% sequence identity to SEQ ID NO: 170, or a functional fragment orvariant thereof.

In some embodiments, the composition of the disclosure comprises anucleic acid molecule or a plasmid comprising at least about 70%, 75%,80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or 100% sequence identity to SEQ ID NO: 161, SEQ ID NO: 162,SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ IDNO: 1676 or SEQ ID NO: 168, or a functional fragment or variant thereof,and an expressible nucleic acid sequence comprising at least about 70%,75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 68, SEQ ID NO: 69,SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO:77, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 83, SEQ IDNO: 84, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 90, SEQID NO: 92, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 98,SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 104, SEQ IDNO: 105, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 111,SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 117, SEQ IDNO: 119, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 125,SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 131, SEQ IDNO: 132, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 138,SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 144, SEQ IDNO: 146, SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 152,SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 158 or SEQ IDNO: 159, or a functional fragment or variant thereof. In someembodiments, the composition of the disclosure comprises a nucleic acidmolecule or a plasmid comprising the nucleotide sequence of SEQ ID NO:161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165, SEQID NO: 166, SEQ ID NO: 1676 or SEQ ID NO: 168, or a functional fragmentor variant thereof, and an expressible nucleic acid sequence comprisingthe nucleotide sequence of SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 71,SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO:78, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 84, SEQ IDNO: 86, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 92, SEQID NO: 93, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 99,SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 105, SEQ IDNO: 107, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 113,SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 119, SEQ IDNO: 120, SEQ ID NO: 122, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 126,SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 132, SEQ IDNO: 134, SEQ ID NO: 135, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 140,SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 146, SEQ IDNO: 147, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 153,SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 158 or SEQ ID NO: 159, or afunctional fragment or variant thereof. In some embodiments, thecomposition of the disclosure comprises a nucleic acid molecule or aplasmid comprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO:164, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 1676 or SEQ ID NO: 168,or a functional fragment or variant thereof, and an expressible nucleicacid sequence encoding a polypeptide comprising at least about 70%, 75%,80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99% or 100% sequence identity to SEQ ID NO: 70, SEQ ID NO: 73, SEQID NO: 76, SEQ ID NO: 79, SEQ ID NO: 82, SEQ ID NO: 85, SEQ ID NO: 88,SEQ ID NO: 91, SEQ ID NO: 94, SEQ ID NO: 97, SEQ ID NO: 100, SEQ ID NO:103, SEQ ID NO: 106, SEQ ID NO: 109, SEQ ID NO: 112, SEQ ID NO: 115, SEQID NO: 118, SEQ ID NO: 121, SEQ ID NO: 124, SEQ ID NO: 127, SEQ ID NO:130, SEQ ID NO: 133, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 142, SEQID NO: 145, SEQ ID NO: 148, SEQ ID NO: 151, SEQ ID NO: 154, SEQ ID NO:157 or SEQ ID NO: 160, or a functional fragment or variant thereof. Insome embodiments, the composition of the disclosure comprises a nucleicacid molecule or a plasmid comprising the nucleotide sequence of SEQ IDNO: 161, SEQ ID NO: 162, SEQ ID NO: 163, SEQ ID NO: 164, SEQ ID NO: 165,SEQ ID NO: 166, SEQ ID NO: 1676 or SEQ ID NO: 168, or a functionalfragment or variant thereof, and an expressible nucleic acid sequenceencoding a polypeptide comprising the amino acid sequence of SEQ ID NO:70, SEQ ID NO: 73, SEQ ID NO: 76, SEQ ID NO: 79, SEQ ID NO: 82, SEQ IDNO: 85, SEQ ID NO: 88, SEQ ID NO: 91, SEQ ID NO: 94, SEQ ID NO: 97, SEQID NO: 100, SEQ ID NO: 103, SEQ ID NO: 106, SEQ ID NO: 109, SEQ ID NO:112, SEQ ID NO: 115, SEQ ID NO: 118, SEQ ID NO: 121, SEQ ID NO: 124, SEQID NO: 127, SEQ ID NO: 130, SEQ ID NO: 133, SEQ ID NO: 136, SEQ ID NO:139, SEQ ID NO: 142, SEQ ID NO: 145, SEQ ID NO: 148, SEQ ID NO: 151, SEQID NO: 154, SEQ ID NO: 157 or SEQ ID NO: 160, or a functional fragmentor variant thereof.

In some embodiments, the disclosure relates to a vector or a plasmidcomprising one or a plurality of regulatory sequences operably linked toone or more of any of the disclosed expressible nucleic acid sequences.In some embodiments, the disclosure relates to a composition comprisinga nucleic acid molecule comprising at least about 70%, 75%, 80%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or100% sequence identity to SEQ ID NO: 161, or a functional fragment orvariant thereof, and positioned within the multiple cloning site thereofis one or more expressible nucleic acid sequences according to thepresent disclosure. In some embodiments, the disclosure relates to acomposition comprising one or a plurality of RNA molecules, eachindividually comprising the RNA sequences disclosed herein, includingbut not limited to SEQ ID NO: 69, SEQ ID NO: 72, SEQ ID NO: 75, SEQ IDNO: 78, SEQ ID NO: 81, SEQ ID NO: 84, SEQ ID NO: 87, SEQ ID NO: 90, SEQID NO: 93, SEQ ID NO: 96, SEQ ID NO: 99, SEQ ID NO: 102, SEQ ID NO: 105,SEQ ID NO: 108, SEQ ID NO: 111, SEQ ID NO: 114, SEQ ID NO: 117, SEQ IDNO: 120, SEQ ID NO: 123, SEQ ID NO: 126, SEQ ID NO: 129, SEQ ID NO: 132,SEQ ID NO: 135, SEQ ID NO: 138, SEQ ID NO: 141, SEQ ID NO: 144, SEQ IDNO: 147, SEQ ID NO: 150, SEQ ID NO: 153, SEQ ID NO: 156 or SEQ ID NO:159, or a functional fragment or variant thereof.

C. Polypeptide Sequences

Disclosed are the polypeptide sequences encoded by the disclosed nucleicacid sequences. In some embodiments, the disclosure relates tocompositions comprising polypeptide sequences encoded by the expressiblenucleic acid molecules of the present disclosure comprising a scaffolddomain comprising a self-assembling polypeptide and an antigen domaincomprising a viral antigen, and optionally comprising a leader domaincomprising a leader sequence and/or a linker domain comprising a linkerpeptide. In some embodiments, the disclosure relates to compositionscomprising polypeptide sequences encoded by the expressible nucleic acidmolecules of the present disclosure comprising a leader domaincomprising a leader sequence and an antigen domain comprising threeviral monomers (trimer), and optionally comprising one or plurality oflinker domains each comprising a linker peptide. The disclosure alsorelates to cells expressing one or more such polypeptides disclosedherein.

In some embodiments, the antigen domain comprised in the polypeptides ofthe disclosure comprises a soluble viral trimer or a soluble monomerthereof, wherein the soluble viral trimer or the soluble monomer thereofis from a virus of the family Coronaviridae. A “viral trimer” refers toa protein complex formed by three covalently or non-convalently boundviral proteins. Each of these viral proteins, when present by itself, iscalled as a “monomer” of the viral trimer.

Solubility of the disclosed viral trimer or monomer thereof can bedetermined using any methods known in the art. Protein solubility is athermodynamic parameter defined as the concentration of protein in asaturated solution that is in equilibrium with a solid phase, eithercrystalline or amorphous, under a given set of conditions. Solubilitycan be influenced by a number of extrinsic and intrinsic factors.Extrinsic factors that influence protein solubility include pH, ionicstrength, temperature, and the presence of various solvent additives.Varying these extrinsic factors can lead to increased solubility;however, altering the solution conditions is not always appropriate orsufficient to increase protein solubility to the extent required. Theintrinsic factors that influence protein solubility are definedprimarily by the amino acids on the protein surface, but a detailedunderstanding of how one can alter the intrinsic properties of a proteinto increase its solubility is lacking.

There are generally two methods used to measure protein solubility inaqueous solution: (1) adding lyophilized protein to solvent; and (2)concentrating a protein solution by ultrafiltration. Both of thesemethods require that the concentration of protein in solution beincreased until saturation is reached; however, this is often difficultto do, especially with very soluble proteins, because gel-like orsupersaturated solutions may form, making it difficult to determine thesolubility values accurately. When lyophilized protein is added tosolvent, the variable water and salt content of the lyophilized powderis difficult to control and can have a significant effect on solubilitymeasurements.

One way to avoid the difficulties of measuring protein solubility is tomake use of an extraneous agent that lowers the solubility of a proteincalled a precipitant. Protein precipitants can be divided into threeclasses: salts, organic solvents, and long-chain polymers. Theseprecipitants are used by crystallographers to achieve slow precipitationand crystal formation; however, they can also be used to induceamorphous precipitation by direct mixing with protein solutions. Commonexamples from each of these three classes of precipitants, respectively,are ammonium sulfate, isopropanol, and polyethylene glycol (PEG). Therelationship between precipitant concentration to protein solubility isdescribed by the following general expression:

Log S=constant−β[Precipitant],  Equation (1)

where S is the measured solubility at a given concentration ofprecipitant, and 13 is the dependence of solubility on precipitantconcentration for a given protein. The constant is the y-intercept ofthe solubility plot, and for PEG precipitations is equal to thelogarithm of the protein activity. For dilute protein solutions, as theactivity constant approaches one, Equation (1) becomes

Log S=Log S _(o)−β[Precipitant],  Equation (2)

where S₀ is the solubility in the absence of precipitant. It is showedthat for PEG precipitations, the linearity of Equation (1) extends tozero precipitant for proteins whose solubility can be accuratelymeasured in buffer alone. In this case, the constant portion of Equation(2), Log S₀, can be used as an estimate of solubility in the absence ofprecipitant. For salts, Equation (1) only describes the salting-outregion of the solubility plot. At low salt concentrations, salting-in isobserved and the solubility is higher than in the absence of salt.Therefore, the constant obtained from salt precipitations represents aprojection of the salting-out region onto the y axis. In someembodiments, the nucleic acid sequences of certain compositions encodeviral monomers that, upon expression, preferentially form into viraltrimers. In some embodiments, the viral antigens form into viral trimerproteins displayed upon a scaffold polypeptide, such as aself-assembling polypeptide polyhedron. In some embodiments, the viralanigens are at a concentration within the disclosed compositions thatdoes not exceed a concentration that causes precipitation out ofsolution. In some embodiments, the scaffold peptide and the displayedviral antigens are soluble in aqueous solution, such as PBS, sterilewater or buffered Ringer's solution.

In some embodiments, the polypeptide encoded by the expressible nucleicacid molecule of the present disclosure comprises at least about 70%,75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 70, SEQ ID NO: 73,SEQ ID NO: 76, SEQ ID NO: 79, SEQ ID NO: 82, SEQ ID NO: 85, SEQ ID NO:88, SEQ ID NO: 91, SEQ ID NO: 94, SEQ ID NO: 97, SEQ ID NO: 100, SEQ IDNO: 103, SEQ ID NO: 106, SEQ ID NO: 109, SEQ ID NO: 112, SEQ ID NO: 115,SEQ ID NO: 118, SEQ ID NO: 121, SEQ ID NO: 124, SEQ ID NO: 127, SEQ IDNO: 130, SEQ ID NO: 133, SEQ ID NO: 136, SEQ ID NO: 139, SEQ ID NO: 142,SEQ ID NO: 145, SEQ ID NO: 148, SEQ ID NO: 151, SEQ ID NO: 154, SEQ IDNO: 157 or SEQ ID NO: 160, or a functional fragment or variant thereof.In some embodiments, the polypeptide is encoded by a nucleic acidsequence comprises at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 72,SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 86, SEQ IDNO: 87, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 93, SEQID NO: 95, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 101,SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 107, SEQ IDNO: 108, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 114,SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, SEQ IDNO: 122, SEQ ID NO: 123, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 128,SEQ ID NO: 129, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 134, SEQ IDNO: 135, SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO: 141,SEQ ID NO: 143, SEQ ID NO: 144, SEQ ID NO: 146, SEQ ID NO: 147, SEQ IDNO: 149, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 155,SEQ ID NO: 156, SEQ ID NO: 158 or SEQ ID NO: 159, or a functionalfragment or variant thereof.

In some embodiments, the leader sequence encoded by the expressiblenucleic acid sequence of the disclosure comprises at least about 70%,75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 5,or a functional fragment or variant thereof. In some embodiments, theleader sequence is encoded by a nucleic acid sequence comprises at leastabout 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 2, SEQID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 6, or a functional fragment orvariant thereof.

In some embodiments, the self-assembling polypeptide encoded by theexpressible nucleic acid sequence of the disclosure comprises at leastabout 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 8, SEQID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18 orSEQ ID NO: 20, or a functional fragment or variant thereof. In someembodiments, the self-assembling polypeptide is encoded by a nucleicacid sequence comprises at least about 70%, 75%, 80%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ IDNO: 13, SEQ ID NO: 15, SEQ ID NO: 17 or SEQ ID NO: 19, or a functionalfragment or variant thereof.

In some embodiments, the linker peptide encoded by the expressiblenucleic acid sequence of the disclosure comprises at least about 70%,75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 22, SEQ ID NO: 23,SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 32, SEQ ID NO:34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ IDNO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQID NO: 54, SEQ ID NO: 56 or SEQ ID NO: 58, or a functional fragment orvariant thereof. In some embodiments, the linker peptide is encoded by anucleic acid sequence comprises at least about 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 21, SEQ ID NO: 27, SEQ ID NO: 28, SEQ IDNO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45,SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO:55 or SEQ ID NO: 57 or a functional fragment or variant thereof.

In some embodiments, the viral antigen or monomer encoded by theexpressible nucleic acid sequence of the disclosure comprises at leastabout 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 60, SEQID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO: 172,SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 or SEQ IDNO: 177, or a functional fragment or variant thereof. In someembodiments, the viral antigen or monomer is encoded by a nucleic acidsequence comprises at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequenceidentity to SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63 or SEQ ID NO:65, or a functional fragment or variant thereof.

In some embodiments, the polypeptides encoded by the expressible nucleicacid molecule of the present disclosure comprises a furin cleavage sitecomprising at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity toSEQ ID NO: 67. In some embodiments, the polypeptides encoded by theexpressible nucleic acid molecule of the present disclosure comprises atransmembrane domain comprising at least about 70%, 75%, 80%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 169. In some embodiments, thepolypeptides encoded by the expressible nucleic acid molecule of thepresent disclosure comprises a foldon domain comprising at least about70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 170.

D. Pharmaceutical Compositions

Disclosed are pharmaceutical compositions comprising any one or more ofthe disclosed compositions and a pharmaceutically acceptable carrier.Disclosed are pharmaceutical compositions comprising therapeuticallyeffective amounts of any one or more of the disclosed compositions and apharmaceutically acceptable carrier.

In some embodiments, any of the disclosed compositions is from about 1to about 30 micrograms of the disclosed DNA and/or RNA vaccine. Forexample, any of the disclosed compositions can be from about 1 to about5 micrograms the disclosed DNA and/or RNA vaccine. In some preferredembodiments, the pharmaceutical compositions contain from about 5nanograms to about 800 micrograms of the disclosed DNA and/or RNAvaccine. In some preferred embodiments, the pharmaceutical compositionscontain about 25 to about 250 micrograms, from about 100 to about 200micrograms, from about 1 nanogram to 100 milligrams; from about 1microgram to about 10 milligrams; from about 0.1 microgram to about 10milligrams; from about 1 milligram to about 2 milligrams, from about 5nanograms to about 1000 micrograms, from about 10 nanograms to about 800micrograms, from about 0.1 to about 500 micrograms, from about 1 toabout 350 micrograms, from about 25 to about 250 micrograms, from about100 to about 200 micrograms of the DNA and/or RNA vaccine or plasmidthereof. The pharmaceutical compositions can comprise from about 5nanograms to about 10 mg of the disclosed DNA and/or RNA vaccine. Insome embodiments, pharmaceutical compositions according to the presentdisclosure comprise from about 25 nanograms to about 5 mg of thedisclosed DNA and/or RNA vaccine. In some embodiments, thepharmaceutical compositions contain from about 50 nanograms to about 1mg of the disclosed DNA and/or RNA vaccine. In some embodiments, thepharmaceutical compositions contain about from about 0.1 to about 500micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions contain from about 1 to about 350micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions contain from about 5 to about 250micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions contain from about 10 to about 200micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions contain from about 15 to about 150micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions contain about 20 to about 100 microgramsof the disclosed DNA and/or RNA vaccine. In some embodiments, thepharmaceutical compositions contain about 25 to about 75 micrograms ofthe disclosed DNA and/or RNA vaccine. In some embodiments, thepharmaceutical compositions contain about 30 to about 50 micrograms ofthe disclosed DNA and/or RNA vaccine. In some embodiments, thepharmaceutical compositions contain about 35 to about 40 micrograms ofthe disclosed DNA and/or RNA vaccine. In some embodiments, thepharmaceutical compositions contain about 100 to about 200 microgramsthe disclosed DNA and/or RNA vaccine. In some embodiments, thepharmaceutical compositions comprise about 10 micrograms to about 100micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions comprise about 20 micrograms to about 80micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions comprise about 25 micrograms to about 60micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions comprise about 30 nanograms to about 50micrograms of the disclosed DNA and/or RNA vaccine. In some embodiments,the pharmaceutical compositions comprise about 35 nanograms to about 45micrograms of the disclosed DNA and/or RNA vaccine. In some preferredembodiments, the pharmaceutical compositions contain about 0.1 to about500 micrograms of the disclosed DNA and/or RNA vaccine. In somepreferred embodiments, the pharmaceutical compositions contain about 1to about 350 micrograms of the disclosed DNA and/or RNA vaccine. In somepreferred embodiments, the pharmaceutical compositions contain about 1to about 250 micrograms of the disclosed DNA and/or RNA vaccine. In somepreferred embodiments, the pharmaceutical compositions contain about 2to about 200 micrograms the disclosed DNA and/or RNA vaccine.

In some embodiments, pharmaceutical compositions according to thepresent disclosure comprise at least about 10, 15, 20, 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100 nanograms of thedisclosed DNA and/or RNA vaccine. In some embodiments, thepharmaceutical compositions can comprise at least about 1, 5, 10, 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95,100, 105,110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175,180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245,250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315,320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385,390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455,460, 465, 470, 475, 480, 485, 490, 495, 500, 605, 610, 615, 620, 625,630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695,700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765,770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835,840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905,910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975,980, 985, 990, 995 or 1000 micrograms of the disclosed DNA and/or RNAvaccine. In some embodiments, the pharmaceutical composition cancomprise at least 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5,8, 8.5, 9, 9.5 or 10 mg or more of the disclosed DNA and/or RNA vaccine.

In other embodiments, the pharmaceutical composition can comprise up toand including about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70,75, 80, 85, 90, 95 or 100 nanograms of the disclosed DNA and/or RNAvaccine. In some embodiments, the pharmaceutical composition cancomprise up to and including about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100,105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170,175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240,245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310,315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380,385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450,455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 605, 610, 615, 620,625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690,695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760,765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830,835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900,905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970,975, 980, 985, 990, 995, or 1000 micrograms of the disclosed DNA and/orRNA vaccine. In some embodiments, the pharmaceutical composition cancomprise up to and including about 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5,6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or about 10 mg of the disclosed DNAand/or RNA vaccine. The pharmaceutical composition can further compriseother agents for formulation purposes according to the mode ofadministration to be used. In cases where pharmaceutical compositionsare injectable pharmaceutical compositions, they are sterile, pyrogenfree and particulate free. An isotonic formulation is preferably used.Generally, additives for isotonicity can include sodium chloride,dextrose, mannitol, sorbitol and lactose. In some cases, isotonicsolutions such as phosphate buffered saline are preferred. Stabilizersinclude gelatin and albumin. In some embodiments, a vasoconstrictionagent is added to the formulation.

The vaccine can further comprise a pharmaceutically acceptableexcipient. The pharmaceutically acceptable excipient can be functionalmolecules as vehicles, adjuvants, carriers, or diluents. Thepharmaceutically acceptable excipient can be a transfection facilitatingagent, which can include surface active agents, such asimmune-stimulating complexes (ISCOMS), Freunds incomplete adjuvant, LPSanalog including monophosphoryl lipid A, muramyl peptides, quinoneanalogs, vesicles such as squalene and squalene, hyaluronic acid,lipids, liposomes, calcium ions, viral proteins, polyanions,polycations, or other known transfection facilitating agents. In someembodiments, the vaccine is a composition comprising a plasmid DNAmolecule, RNA molecule or DNA/RNA hybrid molecule encoding anexpressible nucleic acid sequence, the expressible nucleic acid sequencecomprising a first nucleic acid encoding a self-assembling nanoparticlecomprising a viral antigen, optionally encoding a leader sequencedisclosed herein.

The transfection facilitating agent is a polyanion, polycation,including poly-L-glutamate (LGS), or lipid. The transfectionfacilitating agent is poly-L-glutamate, and more preferably, thepoly-L-glutamate is present in the vaccine at a concentration less than6 mg/ml. The transfection facilitating agent can also include surfaceactive agents such as immune-stimulating complexes (ISCOMS), Freundsincomplete adjuvant, LPS analog including monophosphoryl lipid A,muramyl peptides, quinone analogs and vesicles such as squalene andsqualene, and hyaluronic acid can also be used administered inconjunction with the genetic construct. In some embodiments, the DNAvector vaccines can also include a transfection facilitating agent suchas lipids, liposomes, including lecithin liposomes or other liposomesknown in the art, as a DNA-liposome mixture (see for example WO9324640),calcium ions, viral proteins, polyanions, polycations, or nanoparticles,or other known transfection facilitating agents. In some embodiments,the transfection facilitating agent is a polyanion, polycation,including poly-L-glutamate (LGS), or lipid. Concentration of thetransfection agent in the vaccine is less than about 4 mg/ml, less thanabout 2 mg/ml, less than about 1 mg/ml, less than about 0.750 mg/ml,less than about 0.500 mg/ml, less than about 0.250 mg/ml, less thanabout 0.100 mg/ml, less than about 0.050 mg/ml, or less than about 0.010mg/ml.

The pharmaceutically acceptable excipient can be an adjuvant. Theadjuvant can be other genes that are expressed in alternative plasmid orare deneurological systemed as proteins in combination with the plasmidabove in the vaccine. The adjuvant can be selected from:α-interferon(IFN-α), β-interferon (IFN-β), γ-interferon, plateletderived growth factor (PDGF), TNFα, TNFβ, GM-CSF, epidermal growthfactor (EGF), cutaneous T cell-attracting chemokine (CTACK), epithelialthymus-expressed chemokine (TECK), mucosae-associated epithelialchemokine (MEC), IL-12, IL-15, MHC, CD80, CD86 including IL-15 havingthe signal sequence deleted and optionally including the signal peptidefrom IgE. The adjuvant can be IL-12, IL-15, IL-28, CTACK, TECK, plateletderived growth factor (PDGF), TNFα, TNFβ, GM-CSF, epidermal growthfactor (EGF), IL-1, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-18, or acombination thereof. In an exemplary embodiment, the adjuvant is IL-12.

Other genes which can be useful adjuvants include those encoding: MCP-1,MIP-1a, MIP-1p, IL-8, RANTES, L-selectin, P-selectin, E-selectin, CD34,GlyCAM-1, MadCAM-1, LFA-1, VLA-1, Mac-1, p150.95, PECAM, ICAM-1, ICAM-2,ICAM-3, CD2, LFA-3, M-CSF, G-CSF, IL-4, mutant forms of IL-18, CD40,CD40L, vascular growth factor, fibroblast growth factor, IL-7, nervegrowth factor, vascular endothelial growth factor, Fas, TNF receptor,Fit, Apo-1, p55, WSL-1, DR3, TRAMP, Apo-3, AIR, LARD, NGRF, DR4, DR5,KILLER, TRAIL-R2, TRICK2, DR6, Caspase ICE, Fos, c-jun, Sp-1, Ap-1,Ap-2, p38, p65Rel, MyD88, IRAK, TRAF6, IkB, Inactive NIK, SAP K, SAP-1,JNK, interferon response genes, NFkB, Bax, TRAIL, TRAILrec,TRAILrecDRCS, TRAIL-R3, TRAIL-R4, RANK, RANK LIGAND, Ox40, Ox40 LIGAND,NKG2D, MICA, MICB, NKG2A, NKG2B, NKG2C, NKG2E, NKG2F, TAP1, TAP2 andfunctional fragments thereof or a combination thereof.

In some embodiments adjuvant may be one or more proteins and/or nucleicacid molecules that encode proteins selected from the group consistingof: CCL-20, IL-12, IL-15, IL-28, CTACK, TECK, MEC or RANTES. Examples ofIL-12 constructs and sequences are disclosed in PCT application No.PCT/US1997/019502 (published as WO98/017799) and corresponding U.S.application Ser. No. 08/956,865, and U.S. Provisional Application No.61/569,600 filed Dec. 12, 2011, which are each incorporated herein byreference in their entireties. Examples of IL-15 constructs andsequences are disclosed in PCT application No. PCT/US04/18962 (publishedas WO2005/000235) and corresponding U.S. application Ser. No.10/560,650, and in PCT application No. PCT/US07/00886 (published asWO2007/087178) and corresponding U.S. application Ser. No. 12/160,766,and in PCT Application Serial No. PCT/US10/048827 (published asWO2011/032179), which are each incorporated herein by reference in theirentireties. Examples of IL-28 constructs and sequences are disclosed inPCT application no. PCT/US09/039648 (published as WO2009/124309) andcorresponding U.S. application Ser. No. 12/936,192, which are eachincorporated herein by reference in their entireties. Examples of RANTESand other constructs and sequences are disclosed in PCT application No.PCT/US 1999/004332 (published as WO99/043839) and corresponding U.S.application Ser. No. 09/622,452, which are each incorporated herein byreference in their entieties. Other examples of RANTES constructs andsequences are disclosed in PCT Application No. PCT/US Ser. No.11/024,098 (published as WO2011/097640), which is incorporated herein byreference. Examples of RANTES and other constructs and sequences aredisclosed in PCT Application No. PCT/US 1999/004332 and correspondingU.S. application Ser. No. 09/622,452, which are each incorporated hereinby reference. Other examples of RANTES constructs and sequences aredisclosed in PCT application No. PCT/US11/024098 (published asWO2011/097640), which is incorporated herein by reference in itsentirety. Examples of chemokines CTACK, TECK and MEC constructs andsequences are disclosed in PCT Application No. PCT/US2005/042231(published as WO2007/050095) and corresponding U.S. application Ser. No.11/719,646, which are each incorporated herein by reference in theirentireties. Examples of OX40 and other immunomodulators are disclosed inU.S. application Ser. No. 10/560,653, which is incorporated herein byreference in its entirety. Examples of DR5 and other immunomodulatorsare disclosed in U.S. application Ser. No. 09/622,452, which isincorporated herein by reference in its entirety.

The pharmaceutial compoistion may be formulated according to the mode ofadministration to be used. An injectable vaccine pharmaceuticalcomposition may be sterile, pyrogen free and particulate free. Anisotonic formulation or solution may be used. Additives for isotonicitymay include sodium chloride, dextrose, mannitol, sorbitol, and lactose.The vaccine may comprise a vasoconstriction agent. The isotonicsolutions may include phosphate buffered saline. Vaccine may furthercomprise stabilizers including gelatin and albumin. The stabilizing mayallow the formulation to be stable at room or ambient temperature forextended periods of time such as LGS or polycations or polyanions to thevaccine formulation.

The vaccine can be a DNA or RNA vaccine. In some embodiments, thevaccine is a DNA vaccine. DNA vaccines are disclosed in U.S. Pat. Nos.5,593,972, 5,739,118, 5,817,637, 5,830,876, 5,962,428, 5,981,505,5,580,859, 5,703,055, and 5,676,594, which are incorporated herein fullyby reference. The DNA vaccine can further comprise elements or reagentsthat inhibit it from integrating into the chromosome. Examples ofattenuated live vaccines, those using recombinant vectors to foreignantigens, subunit vaccines and glycoprotein vaccines are described inU.S. Pat. Nos. 4,510,245; 4,797,368; 4,722,848; 4,790,987; 4,920,209;5,017,487; 5,077,044; 5,110,587; 5,112,749; 5,174,993; 5,223,424;5,225,336; 5,240,703; 5,242,829; 5,294,441; 5,294,548; 5,310,668;5,387,744; 5,389,368; 5,424,065; 5,451,499; 5,453,364; 5,462,734;5,470,734; 5,474,935; 5,482,713; 5,591,439; 5,643,579; 5,650,309;5,698,202; 5,955,088; 6,034,298; 6,042,836; 6,156,319 and 6,589,529,which are each incorporated herein by reference in their entireties. Inother embodiments, the vaccine is a RNA vaccine. RNA vaccines aredisclosed in, for example, U.S. Pat. Nos. 10,064,934; 10,064,935;10,124,055; 10,238,731; 10,272,150; 10,383,937; 10,449,244; 10,517,940;10,543,269; 10,675,342; 10,702,597; 10,702,599; and 10,716,846; and U.S.Application Publication Nos. 2011/0033416; 2014/0134129; 2017/0136121;2018/0344838; 2020/0016274; 2020/0030432; 2020/0405844; and2021/0046173, which are each incorporated herein by reference in theirentireties.

In some embodiments, the vaccine is a lipid nanoparticles (LNP)comprising one or a modified RNA molecule. In some embodiments, thevaccine comprises a modified mRNA. Modified polynucleotides (such as,but not limited to, primary constructs), formulations and compositionscomprising modified polynucleotides, and methods of making, using andadministering modified polynucleotides are described in U.S. ProvisionalPatent Application Nos. 61/618,862; 61/681,645; 61/737,130; 61/618,866;61/681,647; 61/737,134; 61/618,868; 61/681,648; 61/737,135; 61/618,873;61/681,650; 61/737,147; 61/618,878; 61/681,654; 61/737,152; 61/618,885;61/681,658; 61/737,155; 61/618,896; 61/668,157; 61/681,661; 61/737,160;61/618,911; 61/681,667; 61/737,168; 61/618,922; 61/681,675; 61/737,174;61/618,935; U61/681,687; 61/737,184; 61/618,945; 61/681,696; 61/737,191;61/618,953; 61/681,704; 61/737,203; 61/681,720; 61/737,213; 61/681,742;61/618,870; 61/681,649; and 61/737,139; and International ApplicationNos. PCT/US2013/030062; PCT/US2013/030064; PCT/US2013/030059;PCT/US2013/030066; PCT/US2013/030067; PCT/US2013/030060;PCT/US2013/030061; PCT/US2013/030070; PCT/US2013/030068;PCT/US2013/030063; and PCT/US2013/031821, the contents of each of whichare herein incorporated by reference in their entireties. Any of therecited polypeptides of the modified polynucleotides of the foregoingare considered useful as a polypeptide of interest or antigen of theLNPs of the present disclosure.

The genetic construct can also be part of a genome of a recombinantviral vector, including recombinant adenovirus, recombinant adenovirusassociated virus and recombinant vaccinia. The genetic construct can bepart of the genetic material in attenuated live microorganisms orrecombinant microbial vectors which live in cells.

In some embodiments, the disclosure relates to a DNA vector pVAX1comprising any one or more of the expressible nucliec acid sequencesdisclosed herein or an RNA transcript thereof. In some embodiments, thedisclosure relates to a pharmaceuical composition comprising a nucleicacid sequencethat includes one or a plurality of the expressible nucleicacid sequences discloed herein or an RNA transcript thereof, and apharmaceutically acceptable carrier.

E. Methods

Provided herein are compositions (e.g., pharmaceutical compositions),methods, kits and reagents for prevention and/or treatment of aninfection of a virus from the family of Coronaviridae, particularly acoronavirus infection, in humans and other mammals. Virus vaccines canbe used as therapeutic or prophylactic agents. They may be used inmedicine to prevent and/or treat infectious disease. In exemplaryaspects, the vaccines of the present disclosure are used to provideprophylactic protection from coronavirus. Prophylactic protection fromcoronavirus can be achieved following administration of aprophylactically effective dose of a coronavirus DNA or RNA vaccine ofthe present disclosure. A prophylactically effective dose is atherapeutically effective dose that prevents infection with the virus ata clinically acceptable level. In some embodiments, the therapeuticallyeffective dose is a dose listed in a package insert for the vaccine.Vaccines can be administered once, twice, three times, four times ormore. It is possible, although less desirable, to administer the vaccineto an infected individual to achieve a therapeutic response. Dosing mayneed to be adjusted accordingly.

In some embodiments, disclosed are methods of vaccinating a subjectcomprising administering a therapeutically effective amount of any ofthe disclosed nucleic acid molecules, compositions, pharmaceuticalcompositions, cells or vaccines to the subject. In some embodiments, thevaccination is against viral infection. In some embodiments, the viralinfection is an infection of a virus from the family of Coronaviridae.In some embodiments, the viral infection is an infection of acoronavirus. In some embodiments, the viral infection is an infection ofSARS-CoV. In some embodiments, the viral infection is an infection ofHCoV NL63. In some embodiments, the viral infection is an infection ofHKU1. In some embodiments, the viral infection is an infection ofMERS-CoV. In some embodiments, the viral infection is an infection ofSARS-CoV-2.

In some embodiments, disclosed are methods of inducing an immuneresponse in a subject comprising administering to the subject any of thedisclosed pharmaceutical compositions. In some embodiments, the methodsare for inducing an immune response against a viral antigen in thesubject. In some embodiments, the immune response induced by thedisclosed methods is against a viral antigen from a virus from thefamily of Coronaviridae. In some embodiments, the immune responseinduced by the disclosed methods is against a viral antigen from acoronavirus. In some embodiments, the immune response induced by thedisclosed methods is against a viral antigen from SARS-CoV. In someembodiments, the immune response induced by the disclosed methods isagainst a viral antigen from HCoV NL63. In some embodiments, the immuneresponse induced by the disclosed methods is against a viral antigenfrom HKU1. In some embodiments, the immune response induced by thedisclosed methods is against a viral antigen from MERS-CoV. In someembodiments, the immune response induced by the disclosed methods isagainst a viral antigen from SARS-CoV-2.

In some embodiments, disclosed are methods of neutralizing one or aplurality of viruses in a subject comprising administering to thesubject any of the disclosed pharmaceutical compositions. In someembodiments, the virus being neutralized by the disclosed method is avirus from the family of Coronaviridae. In some embodiments, the virusbeing neutralized by the disclosed method is a coronavirus. In someembodiments, the virus being neutralized by the disclosed method isSARS-CoV. In some embodiments, the virus being neutralized by thedisclosed method is HCoV NL63. In some embodiments, the virus beingneutralized by the disclosed method is HKU1. In some embodiments, thevirus being neutralized by the disclosed method is MERS-CoV. In someembodiments, the virus being neutralized by the disclosed method isSARS-CoV-2.

In some embodiments, disclosed are methods of neutralizing infection ofone or a plurality of viruses in a subject comprising administering tothe subject any of the disclosed pharmaceutical compositions. In someembodiments, the viral infection being neutralized by the disclosedmethod is an infection of a virus from the family of Coronaviridae. Insome embodiments, the viral infection being neutralized by the disclosedmethod is an infection of coronavirus. In some embodiments, the viralinfection being neutralized by the disclosed method is an infection ofSARS-CoV. In some embodiments, the viral infection being neutralized bythe disclosed method is an infection of HCoV NL63. In some embodiments,the viral infection being neutralized by the disclosed method is aninfection of HKU1. In some embodiments, the viral infection beingneutralized by the disclosed method is an infection of MERS-CoV. In someembodiments, the viral infection being neutralized by the disclosedmethod is an infection of SARS-CoV-2.

In some embodiments, disclosed are methods of stimulating atherapeutically effective antigen-specific immune response against avirus in a mammal infected with the virus comprising administering anyof the disclosed pharmaceutical compositions. In some embodiments, thedisclosed method is a method of stimulating a therapeutically effectiveantigen-specific immune response against a virus from the family ofCoronaviridae. In some embodiments, the disclosed method is a method ofstimulating a therapeutically effective antigen-specific immune responseagainst a coronavirus. In some embodiments, the disclosed method is amethod of stimulating a therapeutically effective antigen-specificimmune response against SARS-CoV. In some embodiments, the disclosedmethod is a method of stimulating a therapeutically effectiveantigen-specific immune response against HCoV NL63. In some embodiments,the disclosed method is a method of stimulating a therapeuticallyeffective antigen-specific immune response against HKU1. In someembodiments, the disclosed method is a method of stimulating atherapeutically effective antigen-specific immune response againstMERS-CoV. In some embodiments, the disclosed method is a method ofstimulating a therapeutically effective antigen-specific immune responseagainst SARS-CoV-2.

In some embodiments, disclosed are methods of inducing expression of aself-assembling vaccine in a subject comprising administering any of thedisclosed pharmaceutical compositions. Also disclosed are methods oftreating a subject having a viral infection or susceptible to becominginfected with a virus comprising administering to the subject atherapeutically effective amount of any of the disclosed pharmaceuticalcompositions. In some embodiments, the viral infection is an infectionof a virus from the family of Coronaviridae. In some embodiments, theviral infection is an infection of coronavirus. In some embodiments, theviral infection is an infection of SARS-CoV. In some embodiments, theviral infection is an infection of HCoV NL63. In some embodiments, theviral infection is an infection of HKU1. In some embodiments, the viralinfection is an infection of MERS-CoV. In some embodiments, the viralinfection is an infection of SARS-CoV-2.

The disclosed pharmaceutical compositions may be administered by anyroute of administration. Accordingly, in some embodiments, theadministration is accomplished by oral administration. In someembodiments, the administration is accomplished by parenteraladministration. In some embodiments, the administration is accomplishedby sublingual administration. In some embodiments, the administration isaccomplished by transdermal administration. In some embodiments, theadministration is accomplished by rectal administration. In someembodiments, the administration is accomplished by transmucosaladministration. In some embodiments, the administration is accomplishedby topical administration. In some embodiments, the administration isaccomplished by inhalation. In some embodiments, the administration isaccomplished by buccal administration. In some embodiments, theadministration is accomplished by intrapleural administration. In someembodiments, the administration is accomplished by intravenousadministration. In some embodiments, the administration is accomplishedby intraarterial administration. In some embodiments, the administrationis accomplished by intraperitoneal administration. In some embodiments,the administration is accomplished by subcutaneous administration. Insome embodiments, the administration is accomplished by intramuscularadministration. In some embodiments, the administration is accomplishedby intranasal administration. In some embodiments, the administration isaccomplished by intrathecal administration. In some embodiments, theadministration is accomplished by intraarticular administration. In someembodiments, the administration is accomplished by intradermaladministration. In some embodiments, the above modes of action areaccomplished by injection of the pharmaceutical compositions disclosedherein.

In some embodiments, the therapeutically effective dose can be fromabout 1 to about 30 micrograms of expressible nucleic acid sequence. Insome embodiments, the therapeutically effective dose can be from about0.001 micrograms of the composition per kilogram of subject to about0.050 micrograms per kilogram of subject.

In some embodiments, any of the disclosed methods can be free ofactivating any mannose-binding lectin or complement process. In someembodiments, any of the disclosed methods is performed without inducingthe MBL-complement pathway.

In some embodiments, the subject can be a human. In some embodiments,the subject is diagnosed with or suspected of having a viral infection.In some embodiments, the subject is diagnosed with or suspected ofhaving an infection of a virus from the family of Coronaviridae. In someembodiments, the subject is diagnosed with or suspected of having aninfection of coronavirus. In some embodiments, the subject is diagnosedwith or suspected of having an infection of SARS-CoV. In someembodiments, the subject is diagnosed with or suspected of having aninfection of HCoV NL63. In some embodiments, the subject is diagnosedwith or suspected of having an infection of HKU1. In some embodiments,the subject is diagnosed with or suspected of having an infection ofMERS-CoV. In some embodiments, the subject is diagnosed with orsuspected of having an infection of SARS-CoV-2.

In some embodiments of the methods of inducing an immune response, theimmune response can be an antigen-specific imune response. In someembodiments, the antigen-specific immune response can be anantigen-specific to SARS-CoV-2 antigen immune response. In someembodiments, the antigen-specific immune response can be atherapeutically effective CD-4+ antigen-specific SARS-CoV-2 immuneresponse. In some embodiments, the antigen-specific immune response canbe a therapeutically effective CD-8+ antigen-specific SARS-CoV-2 immuneresponse. In some embodiments, the antigen-specific immune response canbe a therapeutically effective CD-4+ and CD-8+ antigen-specificSARS-CoV-2 immune response.

In some embodiments, the methods are free of administering anypolypeptide directly to the subject.

In some embodiments, any of the disclosed methods can further compriseadministering to the subject a pharmaceutical composition comprising oneor more pharmaceutically active agents, such as antiviral drugs, amongmany others. In some embodiments, the one or more pharmaceuticallyactive agents include other anticoronarival medications used to inhibitcoronavirus, for example nucleoside analog reverse transcriptaseinhibitors, non-nucleoside reverse transcriptase inhibitors, andprotease inhibitors. Among the available drugs that may be used as apharmaceutically active agent are zidovudine or AZT (or Retrovir®),didanosine or DDI (or Videx®), stavudine or D4T (or Zerit®), lamivudineor 3TC (or Epivir®), zalcitabine or DDC (or Hivid®), abacavir succinate(or Ziagen″), tenofovir disoproxil fumarate salt (or Viread®),emtricitabine (or Emtriva®), Combivir® (contains 3TC and AZT), Trizivir®(contains abacavir, 3TC and AZT); three non-nucleoside reversetranscriptase inhibitors: nevirapine (or Viramune®), delavirdine (orRescriptor®) and efavirenz (or Sustiva®), eight peptidomimetic proteaseinhibitors or approved formulations: saquinavir (or Invirase® orFortovase″), indinavir (or Crixivan®), ritonavir (or Norvir®),nelfinavir (or Viracept″), amprenavir (or Agenerase®), atazanavir(Reyataz), fosamprenavir (or Lexiva), Kaletra®(contains lopinavir andritonavir), and one fusion inhibitor enfuvirtide (or T-20 or Fuzeon®).

In some embodiments, the methods of inducing an immune response caninclude inducing a humoral or cellular immune response. A humoral immuneresponse mainly refers to antibody production. A cellular immuneresponse can include activation of CD4+ T-cells and activation CD8+cells and associated cytotoxic activity. In one aspect, the presentdisclosure features a method of inducing an immune response in asubject, the method comprising administering to the subject in needthereof a pharmaceutically effective amount of any of the nucleic acidmolecules comprising any one or a plurality of the disclosed expressiblenucleic acid sequences or embodiments herein, or any one of thepharmaceutical compositions disclosed herein. In one aspect, the presentdisclosure features a method of inducing a CD8+ T cell immune responsein a subject, the method comprising administering to the subject in needthereof a pharmaceutically effective amount of any of the nucleic acidmolecules comprising any one or a plurality of the disclosed expressiblenucleic acid sequences or embodiments herein, or any one of thepharmaceutical compositions disclosed herein.

In one aspect, the present disclosure features a method of enhancing animmune response in a subject, the method comprising administering to thesubject in need thereof a pharmaceutically effective amount of any ofthe nucleic acid molecules comprising any one or a plurality of thedisclosed expressible nucleic acid sequences or embodiments herein, orany one of the pharmaceutical compositions disclosed herein.

In one aspect, the present disclosure features a method of enhancing aCD8+ T cell immune response in a subject, the method comprisingadministering to the subject in need thereof a pharmaceuticallyeffective amount of any of the nucleic acid molecules comprising any oneor a plurality of the disclosed expressible nucleic acid sequences orembodiments herein, or any one of the pharmaceutical compositionsdisclosed herein.

In some embodiments, the subject has a viral infection and is in need oftherapy for the viral infection. In some embodiments, the viralinfection is an infection of a virus from the family of Coronaviridae.In some embodiments, the viral infection is an infection of coronavirus.In some embodiments, the viral infection is an infection of SARS-CoV. Insome embodiments, the viral infection is an infection of HCoV NL63. Insome embodiments, the viral infection is an infection of HKU1. In someembodiments, the viral infection is an infection of MERS-CoV. In someembodiments, the viral infection is an infection of SARS-CoV-2.

In some embodiments, the subject has previously been treated, and notresponded to anti-viral therapy. In some embodiments, the nucleic acidmolecule and/or the expressible nucleic acid sequence of the disclosureis administered to the subject by electroporation.

The vaccine may be administered by different routes including orally,parenterally, sublingually, transdermally, rectally, transmucosally,topically, via inhalation, via buccal administration, intrapleurally,intravenous, intraarterial, intraperitoneal, subcutaneous,intramuscular, intranasal intrathecal, and intraarticular orcombinations thereof. For veterinary use, the composition may beadministered as a suitably acceptable formulation in accordance withnormal veterinary practice. The veterinarian can readily determine thedosing regimen and route of administration that is most appropriate fora particular animal. The vaccine may be administered by traditionalsyringes, needleless injection devices, “microprojectile bombardmentgone guns,” or other physical methods such as electroporation (“EP”),“hydrodynamic method,” or ultrasound.

The plasmid of the vaccine may be delivered to the mammal by severalwell-known technologies including DNA injection (also referred to as DNAvaccination) with and without in vivo electroporation, liposomemediated, nanoparticle facilitated, recombinant vectors such asrecombinant adenovirus, recombinant adenovirus associated virus andrecombinant vaccinia. The antigen may be delivered via DNA injection andalong with in vivo electroporation.

The vaccine or pharmaceutical composition can be administered byelectroporation. Administration of the vaccine via electroporation ofthe plasmids of the vaccine may be accomplished using electroporationdevices that can be configured to deliver to a desired tissue of amammal a pulse of energy effective to cause reversible pores to form incell membranes, and preferable the pulse of energy is a constant currentsimilar to a preset current input by a user. The electroporation devicemay comprise an electroporation component and an electrode assembly orhandle assembly. The electroporation component may include andincorporate one or more of the various elements of the electroporationdevices, including controller, current waveform generator, impedancetester, waveform logger, input element, status reporting element,communication port, memory component, power source, and power switch.The electroporation can be accomplished using an in vivo electroporationdevice, for example CELLECTRA® EP system (Inovio Pharmaceuticals, Inc.,Blue Bell, Pa.) or Elgen electroporator (Inovio Pharmaceuticals, Inc.)to facilitate transfection of cells by the plasmid.

The electroporation component may function as one element of theelectroporation devices, and the other elements are separate elements(or components) in communication with the electroporation component. Theelectroporation component may function as more than one element of theelectroporation devices, which may be in communication with still otherelements of the electroporation devices separate from theelectroporation component. The elements of the electroporation devicesexisting as parts of one electromechanical or mechanical device may notlimited as the elements can function as one device or as separateelements in communication with one another. The electroporationcomponent may be capable of delivering the pulse of energy that producesthe constant current in the desired tissue, and includes a feedbackmechanism. The electrode assembly may include an electrode array havinga plurality of electrodes in a spatial arrangement, wherein theelectrode assembly receives the pulse of energy from the electroporationcomponent and delivers the same to the desired tissue through theelectrodes. At least one of the plurality of electrodes is neutralduring delivery of the pulse of energy and measures impedance in thedesired tissue and communicates the impedance to the electroporationcomponent. The feedback mechanism may receive the measured impedance andcan adjust the pulse of energy delivered by the electroporationcomponent to maintain the constant current.

A plurality of electrodes may deliver the pulse of energy in adecentralized pattern. The plurality of electrodes may deliver the pulseof energy in the decentralized pattern through the control of theelectrodes under a programmed sequence, and the programmed sequence isinput by a user to the electroporation component. The programmedsequence may comprise a plurality of pulses delivered in sequence,wherein each pulse of the plurality of pulses is delivered by at leasttwo active electrodes with one neutral electrode that measuresimpedance, and wherein a subsequent pulse of the plurality of pulses isdelivered by a different one of at least two active electrodes with oneneutral electrode that measures impedance.

The feedback mechanism may be performed by either hardware or software.The feedback mechanism may be performed by an analog closed-loopcircuit. The feedback occurs every 50 μs, 20 μs, 10 μs or 1 μs, but ispreferably a real-time feedback or instantaneous (i.e., substantiallyinstantaneous as determined by available techniques for determiningresponse time). The neutral electrode may measure the impedance in thedesired tissue and communicates the impedance to the feedback mechanism,and the feedback mechanism responds to the impedance and adjusts thepulse of energy to maintain the constant current at a value similar tothe preset current. The feedback mechanism may maintain the constantcurrent continuously and instantaneously during the delivery of thepulse of energy.

Examples of electroporation devices and electroporation methods that mayfacilitate delivery of the DNA vaccines of the present disclosure,include those described in U.S. Pat. No. 7,245,963 by Draghia-Akli, etal., U.S. Patent Pub. 2005/0052630 submitted by Smith, et al., thecontents of which are hereby incorporated by reference in theirentirety. Other electroporation devices and electroporation methods thatmay be used for facilitating delivery of the DNA vaccines include thoseprovided in co-pending and co-owned U.S. patent application Ser. No.11/874,072, filed Oct. 17, 2007, which claims the benefit under 35 USC119(e) to U.S. Provisional Applications Ser. No. 60/852,149, filed Oct.17, 2006, and 60/978,982, filed Oct. 10, 2007, all of which are herebyincorporated in their entirety.

U.S. Pat. No. 7,245,963 by Draghia-Akli, et al. describes modularelectrode systems and their use for facilitating the introduction of abiomolecule into cells of a selected tissue in a body or plant. Themodular electrode systems may comprise a plurality of needle electrodes;a hypodermic needle; an electrical connector that provides a conductivelink from a programmable constant-current pulse controller to theplurality of needle electrodes; and a power source. An operator cangrasp the plurality of needle electrodes that are mounted on a supportstructure and firmly insert them into the selected tissue in a body orplant. The biomolecules are then delivered via the hypodermic needleinto the selected tissue. The programmable constant-current pulsecontroller is activated and constant-current electrical pulse is appliedto the plurality of needle electrodes. The applied constant-currentelectrical pulse facilitates the introduction of the biomolecule intothe cell between the plurality of electrodes. The entire content of U.S.Pat. No. 7,245,963 is hereby incorporated by reference in its entirety.

U.S. Patent Pub. 2005/0052630 submitted by Smith, et al. describes anelectroporation device which may be used to effectively facilitate theintroduction of a biomolecule into cells of a selected tissue in a bodyor plant. The electroporation device comprises an electro-kinetic device(“EKD device”) whose operation is specified by software or firmware. TheEKD device produces a series of programmable constant-current pulsepatterns between electrodes in an array based on user control and inputof the pulse parameters, and allows the storage and acquisition ofcurrent waveform data. The electroporation device also comprises areplaceable electrode disk having an array of needle electrodes, acentral injection channel for an injection needle, and a removable guidedisk. The entire content of U.S. Patent Pub. 2005/0052630 is herebyincorporated by reference in its entirety. The electrode arrays andmethods described in U.S. Pat. No. 7,245,963 and U.S. Patent Pub.2005/0052630 may be adapted for deep penetration into not only tissuessuch as muscle, but also other tissues or organs. Because of theconfiguration of the electrode array, the injection needle (to deliverthe biomolecule of choice) is also inserted completely into the targetorgan, and the injection is administered perpendicular to the targetissue, in the area that is pre-delineated by the electrodes. Theelectrodes described in U.S. Pat. No. 7,245,963 and U.S. Patent Pub.2005/005263 are preferably 20 mm long and 21 gauge.

Additionally, contemplated in some embodiments that incorporateelectroporation devices and uses thereof, there are electroporationdevices that are those described in U.S. Pat. Nos. 5,273,525; 6,110,161;6,261,281; 6,958,060; and 6,939,862. Furthermore, patents coveringsubject matter provided in U.S. Pat. No. 6,697,669, which concernsdelivery of DNA using any of a variety of devices, and U.S. Pat. No.7,328,064, drawn to a method of injecting DNA, are contemplated herein.The above patents are incorporated by reference in their entireties.

Methods of preparing the nucleic acid molecules are disclosed. In someembodiments, plasmids with one or more multiple cloning sites may bepurchased from commercially available vendors and the expressiblenucleic acids disclosed herein may be ligated into the plasmids after adigestion with a known restriction enzyme needed to cut the plasmid DNA.In another alternative embodiment, membrane-based purification methodsdisclosed herein offer reduced cost, high binding capacity, and highflow rates, resulting in a superior purification process. Thepurification process is further demonstrated to produce plasmid productssubstantially free of genomic DNA, RNA, protein, and endotoxin.

In some embodiments, all of the described aspects of the presentdisclosure are advantageously combined to provide an integrated processfor preparing substantially purified cellular components of interestfrom cells in bioreactors. Again, the cells are most preferablyplasmid-containing cells, and the cellular components of interest aremost preferably plasmids. The substantially purified plasmids aresuitable for various uses, including, but not limited to, gene therapy,plasmid-mediated therapy, as DNA vaccines for human, veterinary, oragricultural use, or for any other application that requires largequantities of purified plasmid. In this aspect, all of the advantagesdescribed for individual aspects of the present disclosure accrue to thecomplete, integrated process, providing a highly advantageous methodthat is rapid, scalable, and inexpensive. Enzymes and otheranimal-derived or biologically sourced products are avoided, as arecarcinogenic, mutagenic, or otherwise toxic substances. Potentiallyflammable, explosive, or toxic organic solvents are similarly avoided.

One aspect of the present disclosure is an apparatus for isolatingplasmid DNA from a suspension of cells having both plasmid DNA andgenomic DNA. An embodiment of the apparatus comprises a first tank andsecond tank in fluid communication with a mixer. The first tank is usedfor holding the suspension cells and the second tank is used for holdinga lysis solution. The suspension of cells from the first tank and thelysis solution from the second tank are both allowed to flow into themixer forming a lysate mixture or lysate fluid. The mixer comprises ahigh shear, low residence-time mixing device with a residence time ofequal to or less than about 1 second. In a preferred embodiment, themixing device comprises a flow through, rotor/stator mixer or emulsifierhaving linear flow rates from about 0.1 L/min to about 20 L/min. Thelysate-mixture flows from the mixer into a holding coil for a period oftime sufficient to lyse the cells and forming a cell lysate suspension,wherein the lysate-mixture has resident time in the holding coil in arange of about 2-8 minutes with a continuous linear flow rate.

The cell lysate suspension is then allowed to flow into a bubble-mixerchamber for precipitation of cellular components from the plasmid DNA.In the bubble mixer chamber, the cell lysate suspension and aprecipitation solution or a neutralization solution from a third tankare mixed together using gas bubbles, which forms a mixed gas suspensioncomprising a precipitate and an unclarified lysate or plasmid containingfluid. The precipitate of the mixed gas suspension is less dense thanthe plasmid containing fluid, which facilitates the separation of theprecipitate from the plasmid containing fluid. The precipitate isremoved from the mixed gas suspension to give a clarified lysate havingthe plasmid DNA, and the precipitate having cellular debris and genomicDNA.

In some embodiments, the bubble mixer-chamber comprises a closedvertical column with a top, a bottom, a first, and a second side with avent proximal to the top of the column. A first inlet port of the bubblemixer-chamber is on the first side proximal to the bottom of the columnand in fluid communication with the holding coil. A second inlet port ofthe bubble mixer-chamber is proximal to the bottom on a second sideopposite of the first inlet port and in fluid communication with a thirdtank, wherein the third tank is used for holding a precipitation or aneutralization solution. A third inlet port of the bubble mixer-chamberis proximal to the bottom of the column and about in the middle of thefirst and second inlets and is in fluid communication with a gas sourcethe third inlet entering the bubble-mixer-chamber. A preferredembodiment utilizes a sintered sparger inside the closed vertical columnof the third inlet port. The outlet port exiting the bubble mixingchamber is proximal to the top of the closed vertical column. The outletport is in fluid communication with a fourth tank, wherein the mixed gassuspension containing the plasmid DNA is allowed to flow from thebubble-mixer-chamber into the fourth tank. The fourth tank is used forseparating the precipitate of the mixed gas suspension having a plasmidcontaining fluid, and can also include an impeller mixer sufficient toprovide uniform mixing of fluid without disturbing the precipitate. Afifth tank is used for a holding the clarified lysate or clarifiedplasmid containing fluid. The clarified lysate is then filtered at leastonce. A first filter has a particle size limit of about 5-10 μm and thesecond filter has a cut of about 0.2 μm. Although gravity, pressure,vacuum, or a mixture thereof can be used for transporting: suspension ofcells; lysis solutions; precipitation solutions; neutralizationsolutions; or mixed gas suspensions from any of the tanks to mixers,holding coils or different tanks, pumps are utilized in a preferredembodiments. In a more preferred embodiment, at least one pump having alinear flow rate from about 0.1 to about 1 ft/second is used.

In another specific embodiment, a Y-connector having a having a firstbifurcated branch, a second bifurcated branch and an exit branch is usedto contact the cell suspension and the lysis solutions before they enterthe high shear, low residence-time mixing device. The first tank holdingthe cell suspension is in fluid communication with the first bifurcatedbranch of the Y-connector through the first pump and the second tankholding the lysis solution is in fluid communication with the secondbifurcated branch of the Y-connector through the second pump. The highshear, low residence-time mixing device is in fluid communication withan exit branch of the Y-connector, wherein the first and second pumpsprovide a linear flow rate of about 0.1 to about 2 ft/second for acontacted fluid exiting the Y-connector.

Another specific aspect of the present disclosure is a method ofsubstantially separating plasmid DNA and genomic DNA from a bacterialcell lysate. The method comprises: delivering a cell lysate into achamber; delivering a precipitation fluid or a neutralization fluid intothe chamber; mixing the cell lysate and the precipitation fluid or aneutralization fluid in the chamber with gas bubbles forming a gas mixedsuspension, wherein the gas mixed suspension comprises the plasmid DNAin a fluid portion (i.e. an unclarified lysate) and the genomic DNA isin a precipitate that is less dense than the fluid portion; floating theprecipitate on top of the fluid portion; removing the fluid portion fromthe precipitate forming a clarified lysate, whereby the plasmid DNA inthe clarified lysate is substantially separated from genomic DNA in theprecipitate. In some embodiments, the chamber is the bubble mixingchamber as described above; the lysing solution comprises an alkali, anacid, a detergent, an organic solvent, an enzyme, a chaotrope, or adenaturant; the precipitation fluid or the neutralization fluidcomprises potassium acetate, ammonium acetate, or a mixture thereof; andthe gas bubbles comprise compressed air or an inert gas. Additionally,the decanted-fluid portion containing the plasmid DNA is preferablyfurther purified with one or more purification steps selected from agroup consisting of: ion exchange, hydrophobic interaction, sizeexclusion, reverse phase purification, endotoxin depletion, affinitypurification, adsorption to silica, glass, or polymeric materials,expanded bed chromatography, mixed mode chromatography, displacementchromatography, hydroxyapatite purification, selective precipitation,aqueous two-phase purification, DNA condensation, thiophilicpurification, ion-pair purification, metal chelate purification,filtration through nitrocellulose, or ultrafiltration.

In some embodiments, a method for isolating a plasmid DNA from cellscomprising: mixing a suspension of cells having the plasmid DNA andgenomic DNA with a lysis solution in ahigh-shear-low-residence-time-mixing-device for a first period of timeforming a cell lysate fluid; incubating the cell lysate fluid for asecond period of time in a holding coil forming a cell lysatesuspension; delivering the cell lysate suspension into a chamber;delivering a precipitation/neutralization fluid into the chamber; mixingthe cell lysate suspension and the a precipitation/neutralization fluidin the chamber with gas bubbles forming a gas mixed suspension, whereinthe gas mixed suspension comprises an unclarified lysate containing theplasmid DNA and a precipitate containing the genomic DNA, wherein theprecipitate is less dense than the unclarified lysate; floating theprecipitate on top of the unclarified lysate; removing the precipitatefrom the unclarified lysate forming a clarified lysate, whereby theplasmid DNA is substantially separated from genomic DNA; precipitatingthe plasmid DNA from the clarified lysate forming a precipitated plasmidDNA; and resuspending the precipitated plasmid DNA in an aqueoussolution.

The disclosure also relates to a method of producing a polypeptide ofinterest in a mammalian cell, the method comprising contacting the cellwith a composition comprising one or a plurality of the RNA moleculesdisclosed herein. In some embodiments, the therapeutic and/orprophylactic agent is an mRNA, and wherein the mRNA encodes thepolypeptide of interest, whereby the mRNA is capable of being translatedin the cell to produce the polypeptide of interest (e.g., nanoparticleor trimer of the disclosure). Compositions comprising RNA nucleic acidsequences of the disclosure can be delivered via lipid-containingnanoparticles and/or modification of the RNA nucleic acid sequenceencoding the one or more viral polypeptides.

In some embodiments, the composition includes at least one RNApolynucleotide having an open reading frame encoding at least oneSARS-CoV-2 antigenic polypeptide having at least one modification, atleast one 5′ terminal cap, and is formulated within a lipidnanoparticle.

In some embodiments, a 5′ terminal cap is 7mG(5′)ppp(5′)NlmpNp. In someembodiments, at least one chemical modification is selected from thegroup consisting of pseudouridine, N1-methylpseudouridine,N1-ethylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine,2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine,2-thio-5-aza-uridine, 2-thio-dihydropseudouridine,2-thio-dihydrouridine, 2-thio-pseudouridine,4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine,4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine,dihydropseudouridine, 5-methoxyuridine, and 2′-O-methyl uridine.

In some embodiments, a lipid nanoparticle comprises a cationic lipid, aPEG-modified lipid, a sterol, and a non-cationic lipid. In someembodiments, a cationic lipid is an ionizable cationic lipid and thenon-cationic lipid is a neutral lipid, and the sterol is a cholesterol.In some embodiments, a cationic lipid is selected from the groupconsisting of 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane(DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate(DLin-MC3-DMA), di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319),(12Z,15Z)-N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine (L608), andN,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecan-8-amine (L530).

In some embodiments, SARS-CoV-2 RNA (e.g., mRNA) vaccines are formulatedin a lipid nanoparticle. In some embodiments, SARS-CoV-2 RNA (e.g.,mRNA) vaccines are formulated in a lipid-polycation complex, referred toas a cationic lipid nanoparticle. The formation of the lipidnanoparticle may be accomplished by methods known in the art and/or asdescribed in U.S. Publication No. 20120178702, herein incorporated byreference in its entirety. As a non-limiting example, the polycation mayinclude a cationic peptide or a polypeptide such as, but not limited to,polylysine, polyornithine and/or polyarginine and the cationic peptidesdescribed in International Publication No. WO2012013326 or U.S.Publication No. US20130142818; each of which is herein incorporated byreference in its entirety. In some embodiments, SARS-CoV-2 RNA (e.g.,mRNA) vaccines are formulated in a lipid nanoparticle that includes anon-cationic lipid such as, but not limited to, cholesterol or dioleoylphosphatidylethanolamine (DOPE).

A lipid nanoparticle formulation may be influenced by, but not limitedto, the selection of the cationic lipid component, the degree ofcationic lipid saturation, the nature of the PEGylation, ratio of allcomponents, and biophysical parameters such as size. In one example bySemple et al. (Nature Biotech. 2010 28:172-176; herein incorporated byreference in its entirety), the lipid nanoparticle formulation iscomposed of 57.1% cationic lipid, 7.1% dipalmitoylphosphatidylcholine,34.3% cholesterol, and 1.4% PEG-c-DMA. As another example, changing thecomposition of the cationic lipid was shown to more effectively deliversiRNA to various antigen presenting cells (Basha et al. Mol Ther. 201119:2186-2200; herein incorporated by reference in its entirety).

In some embodiments, lipid nanoparticle formulations may comprise about35% to about 45% cationic lipid, about 40% to about 50% cationic lipid,about 50% to about 60% cationic lipid and/or about 55% to about 65%cationic lipid. In some embodiments, the ratio of lipid to RNA (e.g.,mRNA) in lipid nanoparticles may be about 5:1 to about 20:1, about 10:1to about 25:1, about 15:1 to about 30:1, and/or at least about 30:1.

In some embodiments, the ratio of PEG in the lipid nanoparticleformulations may be increased or decreased and/or the carbon chainlength of the PEG lipid may be modified from C14 to C18 to alter thepharmacokinetics and/or biodistribution of the lipid nanoparticleformulations. As a non-limiting example, lipid nanoparticle formulationsmay contain about 0.5% to about 3.0%, about 1.0% to about 3.5%, about1.5% to about 4.0%, about 2.0% to about 4.5%, about 2.5% to about 5.0%,and/or about 3.0% to about 6.0% of the lipid molar ratio of PEG-c-DOMG(R-3-[(co-methoxy-poly(ethyleneglycol)2000)carb-amoyl)]-1,2-dimyristyloxypropyl-3-amine) (also referred to hereinas PEG-DOMG) as compared to the cationic lipid, DSPC, and cholesterol.In some embodiments, the PEG-c-DOMG may be replaced with a PEG lipidsuch as, but not limited to, PEG-DSG (1,2-Distearoyl-sn-glycerol,methoxypolyethylene glycol), PEG-DMG (1,2-Dimyristoyl-sn-glycerol)and/or PEG-DPG (1,2-Dipalmitoyl-sn-glycerol, methoxypolyethyleneglycol). The cationic lipid may be selected from any lipid known in theart such as, but not limited to, DLin-MC3-DMA, DLin-DMA, C12-200, andDLin-KC2-DMA.

In some embodiments, a SARS-CoV-2 RNA (e.g., mRNA) vaccine formulationis a nanoparticle that comprises at least one lipid. The lipid may beselected from, but is not limited to, DLin-DMA, DLin-K-DMA, 98N12-5,C12-200, DLin-MC3-DMA, DLin-KC2-DMA, DODMA, PLGA, PEG, PEG-DMG,(12Z,15Z)—N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine (L608),N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecan-8-amine (L530),PEGylated lipids, and amino alcohol lipids.

In some embodiments, a lipid nanoparticle formulation includes about 25%to about 75% on a molar basis of a cationic lipid selected from thegroup consisting of 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane(DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate(DLin-MC3-DMA), and di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), e.g., about35% to about 65%, about 45% to about 65%, about 60%, 57.5%, 50% or 40%on a molar basis.

In some embodiments, a lipid nanoparticle formulation includes about0.5% to about 15% on a molar basis of the neutral lipid, e.g., about 3%to about 12%, about 5% to about 10% or about 15%, 10%, or 7.5% on amolar basis. Examples of neutral lipids include, without limitation,DSPC, POPC, DPPC, DOPE, and SM. In some embodiments, the formulationincludes about 5% to about 50% on a molar basis of the sterol (e.g.,about 15% to about 45%, about 20% to about 40%, about 40%, 38.5%, 35%,or 31% on a molar basis. A non-limiting example of a sterol ischolesterol. In some embodiments, a lipid nanoparticle formulationincludes about 0.5% to about 20% on a molar basis of the PEG orPEG-modified lipid (e.g., about 0.5% to about 10%, about 0.5% to about5%, about 1.5%, 0.5%, 1.5%, 3.5%, or 5% on a molar basis. In someembodiments, a PEG or PEG modified lipid comprises a PEG molecule of anaverage molecular weight of about 2,000 Da. In some embodiments, a PEGor PEG modified lipid comprises a PEG molecule of an average molecularweight of less than about 2,000, for example about 1,500 Da, about 1,000Da, or about 500 Da. Non-limiting examples of PEG-modified lipidsinclude PEG-distearoyl glycerol (PEG-DMG) (also referred herein asPEG-C14 or C14-PEG), and PEG-cDMA (further discussed in Reyes et al. J.Controlled Release, 107, 276-287 (2005) the content of which is hereinincorporated by reference in its entirety).

In some embodiments, lipid nanoparticle formulations include about25-75% of a cationic lipid selected from the group consisting of2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA),dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), anddi((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about0.5-15% of the neutral lipid, about 5-50% of the sterol, and about0.5-20% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include about35-65% of a cationic lipid selected from the group consisting of2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA),dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), anddi((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 3-12%of the neutral lipid, about 15-45% of the sterol, and about 0.5-10% ofthe PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include about45-65% of a cationic lipid selected from the group consisting of2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA),dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), anddi((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 5-10%of the neutral lipid, about 25-40% of the sterol, and about 0.5-10% ofthe PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include about 60%of a cationic lipid selected from the group consisting of2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA),dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), anddi((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 7.5%of the neutral lipid, about 31% of the sterol, and about 1.5% of the PEGor PEG-modified lipid on a molar basis.

Some embodiments of the present disclosure provide a SARS-CoV-2 vaccinethat includes at least one ribonucleic acid (RNA) polynucleotide havingan open reading frame encoding at least one SARS-CoV-2 antigenicpolypeptide, wherein at least about 80% of the uracil in the openreading frame have a chemical modification, optionally wherein theSARS-CoV-2 vaccine is formulated in a lipid nanoparticle. In someembodiments, the RNA vaccine pharmaceutical compositions may beformulated in liposomes such as, but not limited to, DiLa2 liposomes(Marina Biotech, Bothell, Wash.), SMARTICLES® (Marina Biotech, Bothell,Wash.), neutral DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) basedliposomes (e.g., siRNA delivery for ovarian cancer (Landen et al. CancerBiology & Therapy 2006 5(12)1708-1713); herein incorporated by referencein its entirety) and hyaluronan-coated liposomes (Quiet Therapeutics,Israel). In some embodiments, the RNA vaccines may be formulated in alyophilized gel-phase liposomal composition as described in U.S.Publication No. US2012060293, herein incorporated by reference in itsentirety.

The nanoparticle formulations may comprise a phosphate conjugate. Thephosphate conjugate may increase in vivo circulation times and/orincrease the targeted delivery of the nanoparticle. Phosphate conjugatesfor use with the present disclosure may be made by the methods describedin International Publication No. WO2013033438 or U.S. Publication No.US20130196948, the content of each of which is herein incorporated byreference in its entirety. As a non-limiting example, the phosphateconjugates may include a compound of any one of the formulas describedin International Publication No. WO2013033438, herein incorporated byreference in its entirety. In particular, the present disclosure relatesto a pharmaceutical composition comprising nanoparticles which compriseRNA encoding at least one antigen, wherein: (i) the number of positivecharges in the nanoparticles does not exceed the number of negativecharges in the nanoparticles and/or (ii) the nanoparticles have aneutral or net negative charge and/or (iii) the charge ratio of positivecharges to negative charges in the nanoparticles is about 1.4:1 or lessand/or (iv) the zeta potential of the nanoparticles is about 0 or less.

In some embodiments, the nanoparticles described herein are colloidallystable for at least about 2 hours in the sense that no aggregation,precipitation or increase of size and polydispersity index by more thanabout 30% as measured by dynamic light scattering takes place. In someembodiments, the charge ratio of positive charges to negative charges inthe nanoparticles is between about 1.4:1 and about 1:8, preferablybetween about 1.2:1 and about 1:4, e.g. between about 1:1 and about 1:3such as between about 1:1.2 and about 1:2, about 1:1.2 and about 1:1.8,about 1:1.3 and about 1:1.7, in particular between about 1:1.4 and about1:1.6, such as about 1:1.5. In some embodiments, the zeta potential ofthe nanoparticles is about −5 or less, about −10 or less, about −15 orless, about −20 or less or about −25 or less. In various embodiments,the zeta potential of the nanoparticles is about −35 or higher, about−30 or higher or about −25 or higher. In some embodiments, thenanoparticles have a zeta potential from about 0 mV to about −50 mV,preferably about 0 mV to about −40 mV or about −10 mV to about −30 mV.

In some embodiments pharamceutical compositions of the disclosurecomprise a nanoparticle or a liposome that encapsulates a DNA, RNA orDNA/RNA hydbrid comprising at least one expressible nucleic acidsequence. Liposomes are microscopic lipidic vesicles often having one ormore bilayers of a vesicle-forming lipid, such as a phospholipid, andare capable of encapsulating a drug. Different types of liposomes may beemployed in the context of the present disclosure, including, withoutbeing limited thereto, multilamellar vesicles (MLV), small unilamellarvesicles (SUV), large unilamellar vesicles (LUV), sterically stabilizedliposomes (SSL), multivesicular vesicles (MV), and large multivesicularvesicles (LMV) as well as other bilayered forms known in the art. Thesize and lamellarity of the liposome will depend on the manner ofpreparation and the selection of the type of vesicles to be used willdepend on the preferred mode of administration. There are several otherforms of supramolecular organization in which lipids may be present inan aqueous medium, comprising lamellar phases, hexagonal and inversehexagonal phases, cubic phases, micelles, reverse micelles composed ofmonolayers. These phases may also be obtained in the combination withDNA or RNA, and the interaction with RNA and DNA may substantiallyaffect the phase state. The described phases may be present in thenanoparticulate RNA formulations of the present disclosure.

For formation of RNA lipoplexes from RNA and liposomes, any suitablemethod of forming liposomes can be used so long as it provides theenvisaged RNA lipoplexes. Liposomes may be formed using standard methodssuch as the reverse evaporation method (REV), the ethanol injectionmethod, the dehydration-rehydration method (DRV), sonication or othersuitable methods.

After liposome formation, the liposomes can be sized to obtain apopulation of liposomes having a substantially homogeneous size range.

Bilayer-forming lipids have typically two hydrocarbon chains,particularly acyl chains, and a head group, either polar or nonpolar.Bilayer-forming lipids are either composed of naturally-occurring lipidsor of synthetic origin, including the phospholipids, such asphosphatidylcholine, phosphatidylethanolamine, phosphatide acid,phosphatidylinositol, and sphingomyelin, where the two hydrocarbonchains are typically between about 14-22 carbon atoms in length, andhave varying degrees of unsaturation. Other suitable lipids for use inthe composition of the present disclosure include glycolipids andsterols such as cholesterol and its various analogs which can also beused in the liposomes.

Cationic lipids typically have a lipophilic moiety, such as a sterol, anacyl or diacyl chain, and have an overall net positive charge. The headgroup of the lipid typically carries the positive charge. The cationiclipid preferably has a positive charge of about 1 to about 10 valences,more preferably a positive charge of about 1 to about 3 valences, andmore preferably a positive charge of about 1 valence. Examples ofcationic lipids include, but are not limited to1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA);dimethyl-dioctadecylammonium (DDAB);1,2-dioleoyl-3-trimethylammonium-propane (DOTAP);1,2-dioleoyl-3-dimethylammonium-propane (DODAP);1,2-diacyloxy-3-dimethylammonium propanes;1,2-dialkyloxy-3-dimethylammonium propanes; dioctadecyldimethyl ammoniumchloride (DODAC), 1,2-dimyristoyloxypropyl-1,3-dimethylhydroxyethylammonium (DMRIE), and 2,3-dioleoyloxy-N-[2(sperminecarboxamide)ethyl]-N,N-dimethyl-1-propanamium trifluoroacetate (DOSPA).Preferred are DOTMA, DOTAP, DODAC, and DOSPA. Most preferred is DOTMA.

In addition, the nanoparticles described herein preferably furtherinclude a neutral lipid in view of structural stability and the like.The neutral lipid can be appropriately selected in view of the deliveryefficiency of the RNA-lipid complex. Examples of neutral lipids include,but are not limited to,1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (DOPE),1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), diacylphosphatidylcholine, diacylphosphatidyl ethanol amine, ceramide, sphingoemyelin,cephalin, sterol, and cerebroside. Preferred is DOPE and/or DOPC. Mostpreferred is DOPE. In the case where a cationic liposome includes both acationic lipid and a neutral lipid, the molar ratio of the cationiclipid to the neutral lipid can be appropriately determined in view ofstability of the liposome and the like.

According to one embodiment, the nanoparticles described herein maycomprise phospholipids. The phospholipids may be a glycerophospholipid.Examples of glycerophospholipid include, without being limited thereto,three types of lipids: (i) zwitterionic phospholipids, which include,for example, phosphatidylcholine (PC), egg yolk phosphatidylcholine,soybean-derived PC in natural, partially hydrogenated or fullyhydrogenated form, dimyristoyl phosphatidylcholine (DMPC) sphingomyelin(SM); (ii) negatively charged phospholipids: which include, for example,phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidic acid(PA), phosphatidylglycerol (PG) dipalmipoyl PG, dimyristoylphosphatidylglycerol (DMPG); synthetic derivatives in which theconjugate renders a zwitterionic phospholipid negatively charged such isthe case of methoxypolyethylene, glycol-distearoylphosphatidylethanolamine (mPEG-DSPE); and (iii) cationic phospholipids,which include, for example, phosphatidylcholine or sphingomyelin ofwhich the phosphomonoester was O-methylated to form the cationic lipids.

Association of RNA to the lipid carrier can occur, for example, by theRNA filling interstitial spaces of the carrier, such that the carrierphysically entraps the RNA, or by covalent, ionic, or hydrogen bonding,or by means of adsorption by non-specific bonds. Whatever the mode ofassociation, the RNA must retain its therapeutic, i.e. antigen-encoding,properties.

In some embodiments, the nanoparticles comprise at least one lipid. Insome embodiments, the nanoparticles comprise at least one cationiclipid. The cationic lipid can be monocationic or polycationic. Anycationic amphiphilic molecule, eg, a molecule which comprises at leastone hydrophilic and lipophilic moiety is a cationic lipid within themeaning of the present disclosure. In some embodiments, the positivecharges are contributed by the at least one cationic lipid and thenegative charges are contributed by the RNA. In some embodiments, thenanoparticles comprises at least one helper lipid. The helper lipid maybe a neutral or an anionic lipid. The helper lipid may be a naturallipid, such as a phospholipid or an analogue of a natural lipid, or afully synthetic lipid, or lipid-like molecule, with no similarities withnatural lipids. In some embodiments, the cationic lipid and/or thehelper lipid is a bilayer forming lipid.

In some embodiments, the at least one cationic lipid comprises1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA) or analogs orderivatives thereof and/or 1,2-dioleoyl-3-trimethylammonium-propane(DOTAP) or analogs or derivatives thereof. In some embodiments, the atleast one helper lipid comprises1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (DOPE) oranalogs or derivatives thereof, cholesterol (Chol) or analogs orderivatives thereof and/or 1,2-dioleoyl-sn-glycero-3-phosphocholine(DOPC) or analogs or derivatives thereof. In some embodiments, the molarratio of the at least one cationic lipid to the at least one helperlipid is from about 10:0 to about 3:7, preferably about 9:1 to about3:7, about 4:1 to about 1:2, about 4:1 to about 2:3, about 7:3 to about1:1, or about 2:1 to about 1:1, preferably about 1:1. In someembodiments, in this ratio, the molar amount of the cationic lipidresults from the molar amount of the cationic lipid multiplied by thenumber of positive charges in the cationic lipid. In variousembodiments, the lipids are not functionalized such as functionalized bymannose, histidine and/or imidazole, the nanoparticles do not comprise atargeting ligand such as mannose functionalized lipids and/or thenanoparticles do not comprise one or more of the following: pH dependentcompounds, cationic polymers such as polymers containing histidineand/or polylysine, wherein the polymers may optionally be PEGylatedand/or histidylated, or divalent ions such as Ca 2+.

In various embodiments, the RNA nanoparticles may comprise peptides,preferentially with a molecular weight of up to about 2500 Da.

In the nanoparticles described herein the lipid may form a complex withand/or may encapsulate the RNA. In some embodiments, the nanoparticlescomprise a lipoplex or liposome. In some embodiments, the lipid iscomprised in a vesicle encapsulating said RNA. The vesicle may be amultilamellar vesicle, an unilamellar vesicle, or a mixture thereof. Thevesicle may be a liposome. In some embodiments, the nanoparticles arelipoplexes comprising DOTMA and DOPE in a molar ratio of about 10:0 toabout 1:9, preferably about 8:2 to about 3:7, and more preferably ofabout 7:3 to about 5:5 and wherein the charge ratio of positive chargesin DOTMA to negative charges in the RNA is about 1.8:2 to about 0.8:2,more preferably about 1.6:2 to about 1:2, even more preferably about1.4:2 to about 1.1:2 and even more preferably about 1.2:2.

In some embodiments, the nanoparticles are lipoplexes comprising DOTMAand Cholesterol in a molar ratio of about 10:0 to about 1:9, preferablyabout 8:2 to about 3:7, and more preferably of about 7:3 to about 5:5and wherein the charge ratio of positive charges in DOTMA to negativecharges in the RNA is about 1.8:2 to about 0.8:2, more preferably about1.6:2 to about 1:2, even more preferably about 1.4:2 to about 1.1:2 andeven more preferably about 1.2:2. In some embodiments, the nanoparticlesare lipoplexes comprising DOTAP and DOPE in a molar ratio of about 10:0to about 1:9, preferably about 8:2 to about 3:7, and more preferably ofabout 7:3 to about 5:5 and wherein the charge ratio of positive chargesin DOTMA to negative charges in the RNA is about 1.8:2 to about 0.8:2,more preferably about 1.6:2 to about 1:2, even more preferably about1.4:2 to about 1.1:2 and even more preferably about 1.2:2. In someembodiments, the nanoparticles are lipoplexes comprising DOTMA and DOPEin a molar ratio of about 2:1 to about 1:2, preferably about 2:1 toabout 1:1, and wherein the charge ratio of positive charges in DOTMA tonegative charges in the RNA is about 1.4:1 or less. In some embodiments,the nanoparticles are lipoplexes comprising DOTMA and cholesterol in amolar ratio of about 2:1 to about 1:2, preferably about 2:1 to about1:1, and wherein the charge ratio of positive charges in DOTMA tonegative charges in the RNA is about 1.4:1 or less. In some embodiments,the nanoparticles are lipoplexes comprising DOTAP and DOPE in a molarratio of about 2:1 to about 1:2, preferably about 2:1 to about 1:1, andwherein the charge ratio of positive charges in DOTAP to negativecharges in the RNA is about 1.4:1 or less. In some embodiments, thenanoparticles have an avarage diameter in the range of from about 50 nmto about 1000 nm, preferably from about 50 nm to about 400 nm,preferably about 100 nm to about 300 nm such as about 150 nm to about200 nm. In some embodiments, the nanoparticles have a diameter in therange of about 200 to about 700 nm, about 200 to about 600 nm,preferably about 250 to about 550 nm, in particular about 300 to about500 nm or about 200 to about 400 nm.

In some embodiments, the polydispersity index of the nanoparticlesdescribed herein as measured by dynamic light scattering is about 0.5 orless, preferably about 0.4 or less or even more preferably about 0.3 orless. In some embodiments, the nanoparticles described herein areobtainable by one or more of the following: (i) incubation of liposomesin an aqueous phase with the RNA in an aqueous phase, (ii) incubation ofthe lipid dissolved in an organic, water miscible solvent, such asethanol, with the RNA in aqueous solution, (iii) reverse phaseevaporation technique, (iv) freezing and thawing of the product, (v)dehydration and rehydration of the product, (vi) lyophilization andrehydration of the of the product, or (vii) spray drying and rehydrationof the product.

The nanoparticle formulation may comprise a polymer conjugate. Thepolymer conjugate may be a water-soluble conjugate. The polymerconjugate may have a structure as described in U.S. Publication No.20130059360, the content of which is herein incorporated by reference inits entirety. In some aspects, polymer conjugates with thepolynucleotides of the present disclosure may be made using the methodsand/or segmented polymeric reagents described in U.S. Publication No.20130072709, herein incorporated by reference in its entirety. In otheraspects, the polymer conjugate may have pendant side groups comprisingring moieties such as, but not limited to, the polymer conjugatesdescribed in U.S. Publication No. US20130196948, the contents of whichis herein incorporated by reference in its entirety.

The nanoparticle formulations may comprise a conjugate to enhance thedelivery of nanoparticles of the present disclosure in a subject.Further, the conjugate may inhibit phagocytic clearance of thenanoparticles in a subject. In some aspects, the conjugate may be a“self” peptide designed from the human membrane protein CD47 (e.g., the“self” particles described by Rodriguez et al. (Science 2013, 339,971-975), herein incorporated by reference in its entirety). As shown byRodriguez et al., the self peptides delayed macrophage-mediatedclearance of nanoparticles which enhanced delivery of the nanoparticles.In other aspects, the conjugate may be the membrane protein CD47 (e.g.,see Rodriguez et al. Science 2013, 339, 971-975, herein incorporated byreference in its entirety). Rodriguez et al. showed that, similarly to“self” peptides, CD47 can increase the circulating particle ratio in asubject as compared to scrambled peptides and PEG coated nanoparticles.

In some embodiments, about 100% of the uracil in the open reading framehave a chemical modification. In some embodiments, a chemicalmodification is in the 5-position of the uracil. In some embodiments, achemical modification is a N₁-methyl pseudouridine. In some embodiments,about 100% of the uracil in the open reading frame have a N₁-methylpseudouridine in the 5-position of the uracil.

In some embodiments, efficacy of RNA vaccines RNA (e.g., mRNA) can besignificantly enhanced when combined with a flagellin adjuvant, inparticular, when one or more antigen-encoding mRNAs is combined with anmRNA encoding flagellin.

RNA (e.g., mRNA) vaccines combined with the flagellin adjuvant (e.g.,mRNA-encoded flagellin adjuvant) have superior properties in that theymay produce much larger antibody titers and produce responses earlierthan commercially available vaccine formulations. While not wishing tobe bound by theory, it is believed that the RNA vaccines, for example,as mRNA polynucleotides, are better designed to produce the appropriateprotein conformation upon translation, for both the antigen and theadjuvant, as the RNA (e.g., mRNA) vaccines co-opt natural cellularmachinery. Unlike traditional vaccines, which are manufactured ex vivoand may trigger unwanted cellular responses, RNA (e.g., mRNA) vaccinesare presented to the cellular system in a more native fashion.

Some embodiments of the present disclosure provide RNA (e.g., mRNA)vaccines that include at least one RNA (e.g., mRNA) polynucleotidehaving an open reading frame encoding at least one antigenic polypeptideor an immunogenic fragment thereof (e.g., an immunogenic fragmentcapable of inducing an immune response to the antigenic polypeptide) andat least one RNA (e.g., mRNA polynucleotide) having an open readingframe encoding a flagellin adjuvant.

In some embodiments, at least one flagellin polypeptide (e.g., encodedflagellin polypeptide) is a flagellin protein. In some embodiments, atleast one flagellin polypeptide (e.g., encoded flagellin polypeptide) isan immunogenic flagellin fragment. In some embodiments, at least oneflagellin polypeptide and at least one antigenic polypeptide are encodedby a single RNA (e.g., mRNA) polynucleotide. In other embodiments, atleast one flagellin polypeptide and at least one antigenic polypeptideare each encoded by a different RNA polynucleotide.

Some embodiments of the present disclosure provide methods of inducingan antigen specific immune response in a subject, comprisingadministering to the subject a SARS-CoV-2 vaccine in an amount effectiveto produce an antigen specific immune response.

In some aspects, vaccines of the disclosure (e.g., LNP-encapsulated mRNAvaccines) produce prophylactically- and/or therapeutically-efficaciouslevels, concentrations and/or titers of antigen-specific antibodies inthe blood or serum of a vaccinated subject. As defined herein, the termantibody titer refers to the amount of antigen-specific antibodyproduces in s subject, e.g., a human subject. In exemplary embodiments,antibody titer is expressed as the inverse of the greatest dilution (ina serial dilution) that still gives a positive result. In exemplaryembodiments, antibody titer is determined or measured by enzyme-linkedimmunosorbent assay (ELISA). In exemplary embodiments, antibody titer isdetermined or measured by neutralization assay, e.g., bymicroneutralization assay. In certain aspects, antibody titermeasurement is expressed as a ratio, such as about 1:40, 1:100, etc.

In exemplary embodiments of the disclosure, an efficacious vaccineproduces an antibody titer of greater than about 1:40, greater thatabout 1:100, greater than about 1:400, greater than about 1:1000,greater than about 1:2000, greater than about 1:3000, greater than about1:4000, greater than about 1:500, greater than about 1:6000, greaterthan about 1:7500, greater than about 1:10000. In exemplary embodiments,the antibody titer is produced or reached by about 10 days followingvaccination, by about 20 days following vaccination, by about 30 daysfollowing vaccination, by about 40 days following vaccination, or byabout 50 or more days following vaccination. In exemplary embodiments,the titer is produced or reached following a single dose of vaccineadministered to the subject. In other embodiments, the titer is producedor reached following multiple doses, e.g., following a first and asecond dose (e.g., a booster dose.)

In exemplary aspects of the disclosure, antigen-specific antibodies aremeasured in units of g/ml or are measured in units of IU/L(International Units per liter) or mIU/ml (milli International Units perml). In exemplary embodiments of the disclosure, an efficacious vaccineproduces more than about 0.5 μg/ml, 0.1 μg/ml, 0.2 μg/ml, 0.35 μg/ml,0.5 μg/ml, 1 μg/ml, 2 μg/ml, 5 μg/ml or 10 μg/ml of antigen-specificantibodies. In exemplary embodiments of the disclosure, an efficaciousvaccine produces more than about 10 mIU/ml, 20 mIU/ml, 50 mIU/ml, 100mIU/ml, 200 mIU/ml, 500 mIU/ml or 1000 mIU/ml of antigen-specificantibodies. In exemplary embodiments, the antibody level orconcentration is produced or reached by about 10 days followingvaccination, by about 20 days following vaccination, by about 30 daysfollowing vaccination, by about 40 days following vaccination, or byabout 50 or more days following vaccination. In exemplary embodiments,the level or concentration is produced or reached following a singledose of vaccine administered to the subject. In other embodiments, thelevel or concentration is produced or reached following multiple doses,e.g., following a first and a second dose (e.g., a booster dose). Inexemplary embodiments, antibody level or concentration is determined ormeasured by enzyme-linked immunosorbent assay (ELISA). In exemplaryembodiments, antibody level or concentration is determined or measuredby neutralization assay, e.g., by microneutralization assay.

In some embodiments, the SARS-CoV-2 vaccine includes at least one RNApolynucleotide having an open reading frame encoding at least oneSARS-CoV-2 antigenic polypeptide having at least one modification, atleast one 5′ terminal cap, and is formulated within a lipidnanoparticle. 5′-capping of polynucleotides may be completedconcomitantly during the in vitro-transcription reaction using thefollowing chemical RNA cap analogs to generate the 5′-guanosine capstructure according to manufacturer protocols: 3′-O-Me-m7G(5′)ppp(5′) G[the ARCA cap]; G(5′)ppp(5′)A; G(5′)ppp(5′)G; m7G(5′)ppp(5′)A;m7G(5′)ppp(5′)G (New England BioLabs, Ipswich, Mass.). 5′-capping ofmodified RNA may be completed post-transcriptionally using a VacciniaVirus Capping Enzyme to generate the “Cap 0” structure: m7G(5′)ppp(5′)G(New England BioLabs, Ipswich, Mass.). Cap 1 structure may be generatedusing both Vaccinia Virus Capping Enzyme and a 2′-O methyl-transferaseto generate m7G(5′)ppp(5′)G-2′-O-methyl. Cap 2 structure may begenerated from the Cap 1 structure followed by the 2′-O-methylation ofthe 5′-antepenultimate nucleotide using a 2′-O methyl-transferase. Cap 3structure may be generated from the Cap 2 structure followed by the2′-O-methylation of the 5′-preantepenultimate nucleotide using a 2′-Omethyl-transferase. Enzymes are preferably derived from a recombinantsource.

When transfected into mammalian cells, the modified mRNAs have astability of from about 12 to about 18 hours or more than about 18hours, e.g., about 24, 36, 48, 60, 72, or greater than about 72 hours.

In some embodiments, a codon optimized RNA may, for instance, be one inwhich the levels of G/C are enhanced. The G/C-content of nucleic acidmolecules may influence the stability of the RNA. RNA having anincreased amount of guanine (G) and/or cytosine (C) residues may befunctionally more stable than nucleic acids containing a large amount ofadenine (A) and thymine (T) or uracil (U) nucleotides. WO02/098443discloses a pharmaceutical composition containing an mRNA stabilized bysequence modifications in the translated region. Due to the degeneracyof the genetic code, the modifications work by substituting existingcodons for those that promote greater RNA stability without changing theresulting amino acid. The approach is limited to coding regions of theRNA.

In some embodiments, polynucleotides (e.g., RNA polynucleotides, such asmRNA polynucleotides) include a combination of at least two (e.g., 2, 3,4 or more) of the aforementioned modified nucleobases.

In some embodiments, modified nucleobases in polynucleotides (e.g., RNApolynucleotides, such as mRNA polynucleotides) are selected from thegroup consisting of pseudouridine (ψ), 2-thiouridine (s2U),4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine,2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine,2-thio-dihydropseudouridine, 2-thio-dihydrouridine,2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine,4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine,4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine,5-methyluridine, 5-methoxyuridine, 2′-O-methyl uridine,1-methyl-pseudouridine (m1ψ), 1-ethyl-pseudouridine (e1ψ),5-methoxyuridine (mo5U), 5-methyl-cytidine (m5C), α-thio-guanosine,α-thio-adenosine, 5-cyano uridine, 4′-thio uridine 7-deaza-adenine,1-methyl-adenosine (m1A), 2-methyl-adenine (m2A), N6-methyl-adenosine(m6A), and 2,6-Diaminopurine, (I), 1-methyl-inosine (m1I), wyosine(imG), methylwyosine (mimG), 7-deaza-guanosine,7-cyano-7-deaza-guanosine (preQ0), 7-aminomethyl-7-deaza-guanosine(preQ1), 7-methyl-guanosine (m7G), 1-methyl-guanosine (m1G),8-oxo-guanosine, 7-methyl-8-oxo-guanosine, 2,8-dimethyladenosine,2-geranylthiouridine, 2-lysidine, 2-selenouridine,3-(3-amino-3-carboxypropyl)-5,6-dihydrouridine,3-(3-amino-3-carboxypropyl)pseudouridine, 3-methylpseudouridine,5-(carboxyhydroxymethyl)-2′-O-methyluridine methyl ester,5-aminomethyl-2-geranylthiouridine, 5-aminomethyl-2-selenouridine,5-aminomethyluridine, 5-carbamoylhydroxymethyluridine,5-carbamoylmethyl-2-thiouridine, 5-carboxymethyl-2-thiouridine,5-carboxymethyl aminomethyl-2-geranylthiouridine,5-carboxymethylaminomethyl-2-selenouridine, 5-cyanomethyluridine,5-hydroxycytidine, 5-methylaminomethyl-2-geranylthiouridine,7-aminocarboxypropyl-demethylwyosine, 7-aminocarboxypropylwyosine,7-aminocarboxypropylwyosine methyl ester, 8-methyladenosine,N4,N4-dimethylcytidine, N6-formyladenosine, N6-hydroxymethyladenosine,agmatidine, cyclic N6-threonylcarbamoyladenosine, glutamyl-queuosine,methylated undermodified hydroxywybutosine,N4,N4,2′-O-trimethylcytidine, geranylated5-methylaminomethyl-2-thiouridine, geranylated5-carboxymethylaminomethyl-2-thiouridine, Qbase, preQ0base, preQ1base,and combinations of two or more thereof. In some embodiments, the atleast one chemically modified nucleoside is selected from the groupconsisting of pseudouridine, 1-methyl-pseudouridine,1-ethyl-pseudouridine, 5-methylcytosine, 5-methoxyuridine, and acombination thereof. In some embodiments, the polyribonucleotide (e.g.,RNA polyribonucleotide, such as mRNA polyribonucleotide) includes acombination of at least two (e.g., 2, 3, 4 or more) of theaforementioned modified nucleobases. In some embodiments,polynucleotides (e.g., RNA polynucleotides, such as mRNApolynucleotides) include a combination of at least two (e.g., 2, 3, 4 ormore) of the aforementioned modified nucleobases.

The expressible nucleic acid sequence of the present disclosure may bepartially or fully modified along the entire length of the molecule. Forexample, one or more or all or a given type of nucleotide (e.g., purineor pyrimidine, or any one or more or all of A, G, U, C) may be uniformlymodified in a polynucleotide of the disclosure, or in a givenpredetermined sequence region thereof (e.g., in the mRNA including orexcluding the polyA tail). In some embodiments, all nucleotides X in apolynucleotide of the present disclosure (or in a given sequence regionthereof) are modified nucleotides, wherein X may be any one ofnucleotides A, G, U, C, or any one of the combinations A+G, A+U, A+C,G+U, G+C, U+C, A+G+U, A+G+C, G+U+C, or A+G+C.

The polynucleotide may contain from about 1% to about 100% modifiednucleotides (either in relation to overall nucleotide content, or inrelation to one or more types of nucleotide, i.e., any one or more of A,G, U or C) or any intervening percentage (e.g., from about 1% to about20%, from about 1% to about 25%, from about 1% to about 50%, from about1% to about 60%, from about 1% to about 70%, from about 1% to about 80%,from about 1% to about 90%, from about 1% to about 95%, from about 10%to about 20%, from about 10% to about 25%, from about 10% to about 50%,from about 10% to about 60%, from about 10% to about 70%, from about 10%to about 80%, from about 10% to about 90%, from about 10% to about 95%,from about 10% to about 100%, from about 20% to about 25%, from about20% to about 50%, from about 20% to about 60%, from about 20% to about70%, from about 20% to about 80%, from about 20% to about 90%, fromabout 20% to about 95%, from about 20% to about 100%, from about 50% toabout 60%, from about 50% to about 70%, from about 50% to about 80%,from about 50% to about 90%, from about 50% to about 95%, from about 50%to about 100%, from about 70% to about 80%, from about 70% to about 90%,from about 70% to about 95%, from about 70% to about 100%, from about80% to about 90%, from about 80% to about 95%, from about 80% to about100%, from about 90% to about 95%, from about 90% to about 100%, andfrom about 95% to about 100%). It will be understood that any remainingpercentage is accounted for by the presence of unmodified A, G, U, or C.

The nucleic acid sequences may contain at a minimum about 1% and atmaximum about 100% modified nucleotides, or any intervening percentage,such as at least about 5% modified nucleotides, at least about 10%modified nucleotides, at least about 25% modified nucleotides, at leastabout 50% modified nucleotides, at least about 80% modified nucleotides,or at least about 90% modified nucleotides. For example, thepolynucleotides may contain a modified pyrimidine such as a modifieduracil or cytosine. In some embodiments, at least about 5%, at leastabout 10%, at least about 25%, at least about 50%, at least about 80%,at least about 90% or about 100% of the uracil in the polynucleotide isreplaced with a modified uracil (e.g., a 5-substituted uracil). Themodified uracil can be replaced by a compound having a single uniquestructure, or can be replaced by a plurality of compounds havingdifferent structures (e.g., about 2, 3, 4, or more unique structures).In some embodiments, at least about 5%, at least about 10%, at leastabout 25%, at least about 50%, at least about 80%, at least about 90%,or about 100% of the cytosine in the polynucleotide is replaced with amodified cytosine (e.g., a 5-substituted cytosine). The modifiedcytosine can be replaced by a compound having a single unique structure,or can be replaced by a plurality of compounds having differentstructures (e.g., about 2, 3, 4, or more unique structures).

Thus, in some embodiments, the RNA vaccines and/or RNA nucleic acidseqeunces comprise a 5′UTR element, an optionally codon optimized openreading frame, and a 3′UTR element, a poly(A) sequence and/or apolyadenylation signal wherein the RNA is not chemically modified.

Viral vaccines of the present disclosure comprise at least one RNApolynucleotide, such as a mRNA (e.g., modified mRNA). mRNA, for example,is transcribed in vitro from template DNA, referred to as an “in vitrotranscription template.” In some embodiments, the at least one RNApolynucleotide has at least one chemical modification. The at least onechemical modification may include, but is expressly not limited to, anymodification described herein.

In vitro transcription of RNA is known in the art and is described inWO/2014/152027, which is incorporated by reference herein in itsentirety. For example, in some embodiments, the RNA transcript isgenerated using a non-amplified, linearized DNA template in an in vitrotranscription reaction to generate the RNA transcript. In someembodiments, the RNA transcript is capped via enzymatic capping. In someembodiments, the RNA transcript is purified via chromatographic methods,e.g., use of an oligo dT substrate. Some embodiments exclude the use ofDNase. In some embodiments, the RNA transcript is synthesized from anon-amplified, linear DNA template coding for the gene of interest viaan enzymatic in vitro transcription reaction utilizing a T7 phage RNApolymerase and nucleotide triphosphates of the desired chemistry. Anynumber of RNA polymerases or variants may be used in the method of thepresent disclosure. The polymerase may be selected from, but is notlimited to, a phage RNA polymerase, e.g., a T7 RNA polymerase, a T3 RNApolymerase, a SP6 RNa polymerase, and/or mutant polymerases such as, butnot limited to, polymerases able to incorporate modified nucleic acidsand/or modified nucleotides, including chemically modified nucleic acidsand/or nucleotides.

In some embodiments, a non-amplified, linearized plasmid DNA is utilizedas the template DNA for in vitro transcription. In some embodiments, thetemplate DNA is isolated DNA. In some embodiments, the template DNA iscDNA. In some embodiments, the cDNA is formed by reverse transcriptionof a RNA polynucleotide, for example, but not limited to SARS-CoV-2 RNA,e.g. SARS-CoV-2 mRNA. In some embodiments, cells, e.g., bacterial cells,e.g., E. coli, e.g., DH-1 cells are transfected with the plasmid DNAtemplate. In some embodiments, the transfected cells are cultured toreplicate the plasmid DNA which is then isolated and purified. In someembodiments, the DNA template includes a RNA polymerase promoter, e.g.,a T7 promoter located 5′ to and operably linked to the gene of interest.

F. Vaccines

Disclosed are DNA vaccines comprising at least about 70%, 75%, 80%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or100% sequence identity to SEQ ID NO: 68, SEQ ID NO: 71, SEQ ID NO: 74,SEQ ID NO: 77, SEQ ID NO: 80, SEQ ID NO: 83, SEQ ID NO: 86, SEQ ID NO:89, SEQ ID NO: 92, SEQ ID NO: 95, SEQ ID NO: 98, SEQ ID NO: 101, SEQ IDNO: 104, SEQ ID NO: 107, SEQ ID NO: 110, SEQ ID NO: 113, SEQ ID NO: 116,SEQ ID NO: 119, SEQ ID NO: 122, SEQ ID NO: 125, SEQ ID NO: 128, SEQ IDNO: 131, SEQ ID NO: 134, SEQ ID NO: 137, SEQ ID NO: 140, SEQ ID NO: 143,SEQ ID NO: 146, SEQ ID NO: 149, SEQ ID NO: 152, SEQ ID NO: 155 or SEQ IDNO: 158, or a functional fragment or variant thereof. Also disclosed areRNA vaccines comprising at least about 70%, 75%, 80%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%sequence identity to SEQ ID NO: 69, SEQ ID NO: 72, SEQ ID NO: 75, SEQ IDNO: 78, SEQ ID NO: 81, SEQ ID NO: 84, SEQ ID NO: 87, SEQ ID NO: 90, SEQID NO: 93, SEQ ID NO: 96, SEQ ID NO: 99, SEQ ID NO: 102, SEQ ID NO: 105,SEQ ID NO: 108, SEQ ID NO: 111, SEQ ID NO: 114, SEQ ID NO: 117, SEQ IDNO: 120, SEQ ID NO: 123, SEQ ID NO: 126, SEQ ID NO: 129, SEQ ID NO: 132,SEQ ID NO: 135, SEQ ID NO: 138, SEQ ID NO: 141, SEQ ID NO: 144, SEQ IDNO: 147, SEQ ID NO: 150, SEQ ID NO: 153, SEQ ID NO: 156 or SEQ ID NO:159, or a functional fragment or variant thereof. In some embodiment,the DNA or RNA vaccine disclosed herein encodes a polypeptide comprisingat least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ IDNO: 70, SEQ ID NO: 73, SEQ ID NO: 76, SEQ ID NO: 79, SEQ ID NO: 82, SEQID NO: 85, SEQ ID NO: 88, SEQ ID NO: 91, SEQ ID NO: 94, SEQ ID NO: 97,SEQ ID NO: 100, SEQ ID NO: 103, SEQ ID NO: 106, SEQ ID NO: 109, SEQ IDNO: 112, SEQ ID NO: 115, SEQ ID NO: 118, SEQ ID NO: 121, SEQ ID NO: 124,SEQ ID NO: 127, SEQ ID NO: 130, SEQ ID NO: 133, SEQ ID NO: 136, SEQ IDNO: 139, SEQ ID NO: 142, SEQ ID NO: 145, SEQ ID NO: 148, SEQ ID NO: 151,SEQ ID NO: 154, SEQ ID NO: 157 or SEQ ID NO: 160, or a functionalfragment or variant thereof.

The vaccines of the disclosure can be formulated using one or moreexcipients to: increase stability; increase cell transfection; permitthe sustained or delayed release (e.g., from a depot formulation); alterthe biodistribution (e.g., target to specific tissues or cell types);increase the translation of encoded protein in vivo; and/or alter therelease profile of encoded protein (antigen) in vivo. In someembodiments therefore, the disclosed vaccine further comprises apharmaceutically acceptable excipient. In addition to traditionalexcipients such as any and all solvents, dispersion media, diluents, orother liquid vehicles, dispersion or suspension aids, surface activeagents, isotonic agents, thickening or emulsifying agents,preservatives, excipients can include, without limitation, lipidoids,liposomes, lipid nanoparticles, polymers, lipoplexes, core-shellnanoparticles, peptides, proteins, hyaluronidase, nanoparticle mimicsand combinations thereof. In some embodiments, the pharmaceuticallyacceptable excipient is an adjuvant. In some embodiments, the vaccinesof the disclosure do not include an adjuvant (they are adjuvant free).

In some embodiments, the vaccines of the disclosure are formulated in ananoparticle as described herein elsewhere. In some embodiments, thevaccines of the disclosure are formulated in a lipid nanoparticle asdescribed herein elsewhere. In some embodiments, the vaccines of thedisclosure are formulated in a lipid-polycation complex, referred to asa cationic lipid nanoparticle, as described herein elsewhere. As anon-limiting example, the polycation may include a cationic peptide or apolypeptide such as, but not limited to, polylysine, polyornithineand/or polyarginine. In some embodiments, the vaccines of the disclosureare formulated in a lipid nanoparticle that includes a non-cationiclipid such as, but not limited to, cholesterol or dioleoylphosphatidylethanolamine (DOPE) as described herein elsewhere.

In some embodiments, the vaccines of the disclosure are administeredprophylactically as part of an active immunization scheme to healthyindividuals. In some embodiments, the vaccines of the disclosure areadministered therapeutically early in infection during the incubationphase. In some embodiments, the vaccines of the disclosure areadministered therapeutically during active infection after onset ofsymptoms. In some embodiments, the amount of DNA or RNA (e.g., mRNA)vaccine of the present disclosure provided to a cell, a tissue or asubject may be an amount effective for immune prophylaxis. In someembodiments, the vaccines of the disclosure are administrated with otherprophylactic or therapeutic compounds. In some embodiments, theprophylactic or therapeutic compound is an adjuvant. In someembodiments, the prophylactic or therapeutic compound is a booster. Asused herein, when referring to a prophylactic composition, such as avaccine, the term “booster” refers to an extra administration of theprophylactic (vaccine) composition. A booster (or booster vaccine) maybe given after an earlier administration of the prophylacticcomposition. The time of administration between the initialadministration of the prophylactic composition and the booster may be,but is not limited to, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25,30, 35, 40, 45, 50 or 55 minutes, or about 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 or 23 hours, or about1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or 30 days, or about 1, 2, 3,4, 5, or 6 weeks, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12months, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40,45 or 50 years. In some embodiments, the time of administration betweenthe initial administration of the prophylactic composition and thebooster is about 1 week. In some embodiments, the time of administrationbetween the initial administration of the prophylactic composition andthe booster is about 2 weeks. In some embodiments, the time ofadministration between the initial administration of the prophylacticcomposition and the booster is about 3 weeks. In some embodiments, thetime of administration between the initial administration of theprophylactic composition and the booster is about 1 month. In someembodiments, the time of administration between the initialadministration of the prophylactic composition and the booster is about2 months. In some embodiments, the time of administration between theinitial administration of the prophylactic composition and the boosteris about 3 months. In some embodiments, the time of administrationbetween the initial administration of the prophylactic composition andthe booster is about 6 months. In some embodiments, the time ofadministration between the initial administration of the prophylacticcomposition and the booster is about 1 year.

G. Kits

The materials described above as well as other materials can be packagedtogether in any suitable combination as a kit useful for performing, oraiding in the performance of, the disclosed method. It is useful if thekit components in a given kit are designed and adapted for use togetherin the disclosed method. For example disclosed are kits comprising anyof the elements of the disclosed nucleic acid compositions. For example,disclosed are kits comprising nucleic acid sequences comprising a leadersequence, a linker sequence, a nucleic acid sequence encoding aself-assembling polypeptide, and/or a nucleic acid sequence encoding aviral antigen. In some embodiments, the kits can further comprise aplasmid backbone.

Other embodiments are described in the following non-limiting Examples.Various publications, including patents, published applications,technical articles and scholarly articles are cited throughout thespecification. Each of these cited publications is incorporated byreference herein in its entirety.

EXAMPLES Example 1. Experimental Validation of Prefusion CoV2 Trimers

The SARS-CoV-2 prefusion trimers would be tested by size-exclusionchromatography, multi-angle light scattering, ELISA binding to variousreceptors and anti-CoV2 antibodies, negative-stain EM to determinerelative populations of prefusion states, SPR binding to variousreceptors and anti-CoV2 antibodies, native page western blots andthermal stability in buffer and serum using DSF. Collectively, thesemeasurements will enable us to determine if the SARS-CoV-2 trimers havethe relevant design characteristics. The trimers must demonstrate ahomogenous, trimeric oligomerization state as determined by SEC-MALS,western blot and negative stain EM. The trimers must also havealterations in frequencies of relevant prefusion conformations asdetermined by negative stain EM. The trimers must bind to receptors(such as ACE2) or selected anti-CoV2 antibodies as determined by ELISAand SPR. The trimers must have an increase in thermal stability in PBSand serum as determined by DSF. Other equivalent biophysical methods todetermine these parameters may be alternatively employed.

Size-Exclusion Chromatography and Multi-Angle Light Scattering

Expi293F cells will be transfected with a plasmid vector carrying thetrimer transgene with PEI/OPTI-MEM and harvested 6 dayspost-transfection. Transfection supernatant will be first purified witheither affinity chromatography using the AKTA pure 25 system and an IMACNickel column OR gravity flow columns filled with GNL Lectin beads. Theeluate fractions from the affinity purification will be pooled,concentrated and dialyzed into 1×PBS buffer before being loaded onto theSEC column and then purified with size exclusion chromatography, forwhich the Superdex 200 Increase 10/300 GL column or Superose 6 Increase10/300 GL column or equivalent will be run at 0.5 mL/min. Identifiedeluate fractions will then collected and concentrated to 1 mg/mL in PBS.The oligomeric state of the trimers will then be confirmed by sizeexclusion chromatography-multi-angle light scattering (SECMALS) usingthe DAWN HELEOS II multi-angle light scattering system with OptilabT-rEX refractometer (Wyatt Technology).

ELISA

The trimers will be assessed by ELISAs to determine binding againsthuman ACE2 with a detectable tag. Nunc ELISA plates will be coated with1 μg/ml rabbit anti-His6X in 1× PBS for 4-6 hours at room temperatureand washed 4 times with washing buffer (1× PBS and 0.05% Tween 20).Plates will be blocked overnight at 4° C. with blocking buffer (1× PBS,0.05% Tween 20, 5% evaporated milk and 1% FBS). Plates will be washedfour times with washing buffer then incubated with full length (S1+S2)spike protein containing a C-terminal His tag (Sino Biologics, cat.40589-V08B1) at 10 ug/ml for 1 hour at room temperature. Plates will bewashed and then serial dilutions of recombinant human ACE2 with a humanFc tag (ACE2-IgHu) and incubated for 1-2 hours at room temperature.Plates will be again washed and then incubated with 1:10,000 dilution ofhorseradish peroxidase (HRP) conjugated anti-human IgG secondaryantibody (Bethyl, cat. A80-304P) and incubated for 1 hour at roomtemperature. After final wash plates will be developed using 1-StepUltra TMB-ELISA Substrate (ThermoFisher, cat. 34029) and the reactionstopped with 1 M Sulfuric Acid. Plates will be read at 450 nm wavelengthwithin 30 minutes using a SpectraMax Plus 384 Microplate Reader(Molecular Devices, Sunnyvale, Calif.).

Negative Stain EM of Purified SARS-CoV-2 Trimers

The trimers will be produced in Expi293 cells, purified using Agarosebound lectin beads (Agarose Galanthus Nivalis Lectin, VectorLaboratories) followed by size exclusion chromatography (GE Healthcare)using the Superose 6 Increase 10/300 GL column. The proteins will befurther dialyzed into Tris-buffered saline (TBS). A total of 3 μL ofpurified proteins was adsorbed onto glow discharged carbon-coated Cu400EM grids. The grids will be then stained with 3 μL of 2% uranyl acetate,blotted, and stained again with 3 μL of the stain followed by a finalblot. Image collection and data processing will be performed on a FEITecnai T12 microscope equipped with a Oneview Gatan camera at 90,450×magnification at the camera and a pixel size of 1.66 Å.

Surface Plasmon Resonance (SPR)

Kinetics and affinities of antibody-antigen interactions will bemeasured on a Biacore 8K (GE/Cytiva) using CAP (GE/Cytiva) or Protein A(GE/Cytiva) chips and 1× HBS-EP+ pH 7.4 running buffer (20× stock fromTeknova, Cat. No H8022) supplemented with BSA at 1 mg/ml. Typically,100-200 RUs of antigen or antibody will be captured onto each flow celland analytes will be passed over the flow cell at 50 μL/min for 3 minfollowed by a 5 min dissociation time. Regeneration is accomplishedusing 6M Guanidine-HCL in 02.5M NaOH or 10 mM glycine-HCl at pH 1.5 with180 seconds contact time. Raw sensograms will be analyzed using Biacoreevaluation software (GE/Cytiva), including flow cell and blank doublereferencing, and either Equilibrium fits or Kinetic fits with Langmuirmodel, or both, will be employed when applicable. Analyte concentrationswill be measured on a NanoDrop 2000c Spectrophotometer using Absorptionsignal at 280 nm.

Differential Scanning Fluorimetry

The assay will employ a real-time PCR instrument to monitor changes influorescence of SYPRO Orange (or similar dye) and to determine meltingtemperatures. At least three temperature gradient scans will be run fromroom temperature to 120C at rate of 1-2C/min. Signals are normalizedfrom 0 to 100. The SYPRO orange dye and the trimers added at appropriateconcentration (2×-10× range and 100-1000 ug/ml, respectively) tooptimize the experimental conditions. SYPRO orange has an excitationmaximum near 500 nm and emission maximum near 600 nm. The real timesystem channel will be selected to optimize excitation and emission ofSPYRO orange.

Western Blot

Proteins will be separated on a 4-12% BIS-TRIS gel (ThermoFisherScientific), then following transfer, blots will be incubated with ACE2or an anti-SARS-CoV spike protein polyclonal antibody (NovusBiologicals) then visualized with horseradish peroxidase(HRP)-conjugated anti-mouse IgG (GE Amersham).

Immunofluorescence of Transfected 293T Cells

For in vitro staining of trimer protein expression 293T cells will becultured on 4-well glass slides (Lab-Tek) and transfected with 3 μg perwell of pDNA using TurboFectin8.0 (OriGene) transfection reagentfollowing the manufacturer's protocol. Cells will be fixed 48 hrs aftertransfection with 10% Neutral-buffered Formalin (BBC Biochemical, Wash.State) for 10 min at room temperature (RT) and then washed with PBS.Before staining, chamber slides will be blocked with 0.3% (v/v) Triton-X(Sigma), 2% (v/v) donkey serum in PBS for 1 hr at RT. Cells will bestained with a rabbit anti-SARS-CoV spike protein polyclonal antibody(Novus Biologicals) diluted in 1% (w/v) BSA (Sigma), 2% (v/v) donkeyserum, 0.3% (v/v) Triton-X (Sigma) and 0.025% (v/v) 1 g ml⁻¹ SodiumAzide (Sigma) in PBS for 2 hrs at RT. Slides will be washed three timesfor 5 min in PBS and then stained with donkey anti-rabbit IgG AF488(lifetechnologies) for 1 hr at RT. Slides will be washed again andmounted and covered with DAPI-Fluoromount (SouthernBiotech).

Example 2. Vaccine Delivery Systems Recombinant Protein Vaccines

The SARS-CoV-2 Spike trimers described herein are amenable to deliveryas a recombinant subunit vaccine. The trimers can be encoded intomammalian expression plasmids, such as pHLSEC. These plasmids can beemployed to expressed and purified recombinant vaccines as described inthis document. The recombinant proteins can be easily formulated withany number of adjuvant systems (such as aluminum phosphate) to enhanceimmunity.

DNA Plasmid Vaccines

The SARS-CoV-2 Spike trimers described herein are amenable to deliveryas a transgene in a DNA plasmid (such as Inovio Pharmaceutical's synDNAplatform). DNA vaccines are non-infectious and have a long record ofsafety in humans. DNA vaccines can induce high seroconversion rates inpeople. DNA vaccines can induce both B and T cell immunities. DNAplasmids can be formulated with adjuvating material to improve take-up,immune trafficking and resulting immune responses.

RNA Vaccines

The SARS-CoV-2 Spike trimers described herein are amenable to deliveryas a transgene in an RNA vaccine (such as Moderna's mRNA platform). RNAvaccines are non-infectious, non-integrating and are naturallydegrading. RNA vaccines have been shown to stimulate strong B cellresponses to encoded transgenes. Naked RNA can be formulated with lipidnanoparticles (LNPs) to encapsulate RNA protecting the transgene fromdegradation.

Self-Replicating RNA Vaccines

The SARS-CoV-2 Spike trimers described herein are amenable to deliveryas a transgene in a self-replicating RNA vaccine formulated as DNA. RNAreplicons can be delivered by a DNA or mRNA to self-amplify resulting inhigher transgene expression.

Adeno-Associated Virus Vaccines:

Modified versions of the SARS-CoV-2 Spike trimers described herein (suchas ones which are capable of cell fusion by removal of key mutationsimpacting this function) are amenable to delivery as a transgene in AAV(or similar vector). AAV can be employed to obtain high expression ofvaccine antigens in vivo. Numerous AAV variants exist to help targetspecific cell types and each can be explored with a SARS-CoV-2 Spiketrimer.

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1. A composition comprising an expressible nucleic acid sequencecomprising: a) a first nucleic acid sequence encoding a scaffold domaincomprising a self-assembling polypeptide; and b) a second nucleic acidsequence encoding a viral antigen from a virus of the familyCoronaviridae.
 2. The composition of claim 1, wherein theself-assembling polypeptide is from Aquifex aeolicus, Helicobacterpylori, Pyrococcus furiosus or Thermotoga maritima.
 3. The compositionof claim 1, wherein the self-assembling polypeptide comprises at least70% sequence identity to SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, or a functionalfragment or variant thereof.
 4. (canceled)
 5. The composition of claim1, wherein the viral antigen is an antigen from SARS-CoV-2 comprising atleast 70% sequence identity to SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO:64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176 or SEQ ID NO: 177, or afunctional fragment or variant thereof.
 6. (canceled)
 7. The compositionof claim 1, wherein the expressible nucleic acid sequence furthercomprises a third nucleic acid sequence encoding a leader sequence. 8.(canceled)
 9. The composition of claim 1, wherein the expressiblenucleic acid sequence further comprises a nucleic acid sequence encodinga linker, said nucleic acid sequence positioned between the firstnucleic acid sequence and the second nucleic acid sequence in the 5′ to3′ orientation.
 10. (canceled)
 11. The composition of claim 1, whereinthe expressible nucleic acid sequence comprises at least 70% sequenceidentity to SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 72,SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO:80, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 86, SEQ IDNO: 87, SEQ ID NO: 89 or SEQ ID NO: 90, or a functional fragment orvariant thereof; wherein the expressible nucleic acid sequence encodes apolypeptide comprising at least 70% sequence identity to SEQ ID NO: 70,SEQ ID NO: 73, SEQ ID NO: 76, SEQ ID NO: 79, SEQ ID NO: 82, SEQ ID NO:85, SEQ ID NO: 88 or SEQ ID NO: 91, or a functional fragment or variantthereof.
 12. (canceled)
 13. A composition comprising an expressiblenucleic acid sequence comprising: a) a first nucleic acid sequenceencoding a leader sequence; and b) a second nucleic acid sequenceencoding a soluble viral trimer or a soluble monomer thereof, whereinthe soluble viral trimer or the soluble monomer thereof is from a virusof the family Coronaviridae.
 14. The composition of claim 13, whereinthe leader sequence comprises at least 70% sequence identity to SEQ IDNO: 1 or SEQ ID NO: 5, or a functional fragment or variant thereof. 15.The composition of claim 13, wherein the soluble viral trimer or thesoluble monomer thereof is from a coronavirus.
 16. (canceled)
 17. Thecomposition of claim 13, wherein the soluble viral trimer or the solublemonomer thereof comprises at least 70% sequence identity to SEQ ID NO:60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 171, SEQ IDNO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176or SEQ ID NO: 177, or a functional fragment or variant thereof.
 18. Thecomposition of claim 13, wherein the expressible nucleic acid sequencefurther comprises a third nucleic acid sequence encoding a linker. 19.(canceled)
 20. The composition of claim 13, wherein the soluble viraltrimer or the soluble monomer thereof comprises at least a portion ofSARS-CoV-2 spike protein.
 21. The composition of claim 13, wherein theexpressible nucleic acid sequence further comprises a nucleic acidsequence encoding a self-assembling polypeptide or a functional fragmentor variant thereof.
 22. (canceled)
 23. The composition of claim 1,wherein the expressible nucleic acid sequence comprises at least 70%sequence identity to SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 95, SEQ IDNO: 96, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 102,SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 108, SEQ IDNO: 110, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 116,SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 122, SEQ IDNO: 123, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 128, SEQ ID NO: 129,SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 134, SEQ ID NO: 135, SEQ IDNO: 137, SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 143,SEQ ID NO: 144, SEQ ID NO: 146, SEQ ID NO: 147, SEQ ID NO: 149, SEQ IDNO: 150, SEQ ID NO: 152, SEQ ID NO: 153, SEQ ID NO: 155, SEQ ID NO: 156,SEQ ID NO: 158 or SEQ ID NO: 159, or a functional fragment or variantthereof or wherein the expressible nucleic acid sequence encodes apolypeptide comprising at least 70% sequence identity to SEQ ID NO: 94,SEQ ID NO: 97, SEQ ID NO: 100, SEQ ID NO: 103, SEQ ID NO: 106, SEQ IDNO: 109, SEQ ID NO: 112, SEQ ID NO: 115, SEQ ID NO: 118, SEQ ID NO: 121,SEQ ID NO: 124, SEQ ID NO: 127, SEQ ID NO: 130, SEQ ID NO: 133, SEQ IDNO: 136, SEQ ID NO: 139, SEQ ID NO: 142, SEQ ID NO: 145, SEQ ID NO: 148,SEQ ID NO: 151, SEQ ID NO: 154, SEQ ID NO: 157 or SEQ ID NO: 160, or afunctional fragment or variant thereof. 24.-27. (canceled)
 28. Apharmaceutical composition comprising (i) the composition of claim 13,and (ii) a pharmaceutically acceptable carrier.
 29. The pharmaceuticalcomposition of claim 28, wherein the composition of claim 13 is in anamount from about 1 to about 100 micrograms.
 30. (canceled)
 31. A methodof vaccinating a subject comprising administering a therapeuticallyeffective amount of the pharmaceutical composition of claim 28 to thesubject. 32.-34. (canceled)
 35. The method of claim 31, wherein themethod is free of activating any mannose-binding lectin or complementprocess. 36.-37. (canceled)
 38. The method of claim 31, wherein thetherapeutically effective dose is from about 0.001 microgram ofcomposition per kilogram of the subject to about 0.05 microgram perkilogram of the subject.
 39. A method of inducing an immune response ina subject comprising administering to the subject the pharmaceuticalcomposition of claim
 28. 40.-50. (canceled)
 51. A method of neutralizingone or a plurality of viruses in a subject comprising administering tothe subject the pharmaceutical composition of claim
 28. 52.-67.(canceled)
 68. A method of inducing expression of a self-assemblingvaccine in a subject comprising administering the pharmaceuticalcomposition of claim
 28. 69. The method of claim 68, wherein the methodis free of administering any polypeptide directly to the subject.70.-126. (canceled)